MXPA00005709A

MXPA00005709A - Bone morphogenetic protein (bmp)-17 and bmp-18 compositions

Info

Publication number: MXPA00005709A
Application number: MXPA/A/2000/005709A
Authority: MX
Inventors: Anthony J Celeste; Beth L Murray
Original assignee: Genetics Institute Inc
Priority date: 1997-12-10
Filing date: 2000-06-09
Publication date: 2001-12-04

Abstract

Purified BMP-17 and BMP-18 proteins and processes for producing them are disclosed. DNA molecules encoding the BMP-17 and BMP-18 proteins are also disclosed. The proteins may be used in the treatment of bone, cartilage, other connective tissue defects and disorders, including tendon, ligament and meniscus, in wound healing and related tissue repair, as well as for treatment of disorders and defects to tissues which include epidermis, nerve, muscle, including cardiac muscle, and other tissues and wounds, and organs such as liver, lung, epithelium, brain, spleen, cardiac, pancreas and kidney tissue. The proteins may also be useful for the induction of growth and/or differentiation of undifferentiated embryonic and stem cells.

Description

COMPOSITIONS OF MORPHOGENETIC PROTEINS OF BONES (PMH) -17 AND PMH-18 FIELD OF THE INVENTION The present invention relates to a novel family of purified proteins, designated as Proteins, Morphogenetic of Bones (PMH) -17 and PMH-18, to related proteins, and to the DNA that encodes them, and to processes to obtain them. These proteins can be used to induce the formation of bone and / or cartilage or other connective tissue, and in wound healing and tissue repair. These proteins can also be used to increase the activity of other bone morphogenetic proteins.

BACKGROUND OF THE INVENTION The search for the molecule or molecules responsible for the inducing activity of bones, cartilage and other connective tissues, present in extracts of bones and other tissues, has led to the discovery of a novel group of molecules called Morphogenetic Proteins of Bones (PHM). The structures of several proteins, designated from PMH-1 to PMH-16 have been elucidated previously. The only inducing activities of these proteins, together with their presence in the bones, suggest that they are important regulators of bone repair processes, and may be involved in the normal maintenance of bone tissue. There is a need to identify if there are additional proteins, particularly human proteins, that play a role in these processes. The present invention relates to the identification of this novel human protein, to which the inventors have designated PMH-17 and PMH-18. Human PMH-17 and PMH-18 appear to be human homologs of a murine protein called Lefty. The nucleotide and amino acid sequences of murine Lefty are described in Zhou et al., Nature, 361: 543-547 (1993). It has been described that the murine Lefty gene is expressed in the mouse node during gastrulation. A human protein, related, designated factor associated with endometrial bleeding [EBAF] was published in Kothapelli et al., J. Clin. Invest. , 99: 2342-2350 (1997).

SUMMARY OF THE INVENTION As used herein, the term "PMH-17 and PMH-18 proteins" refers to the human PMH-17 and PMH-18 proteins, which have the amino acid sequences specified in SEQUENCE (SEQ) ID No.:2 and SEQ ID No.:4, as well as the DNA sequences encoding the PMH-17 and PMH-18 proteins, such as the native human sequences shown in SEQ ID No.:ly and SEQ ID No .: 3. Also included are the natural allelic sequences of SEQ ID No.: SEQ ID No.:3, and codon sequences, degenerate, equivalent, of the above. The DNA sequences of PMH-17 (SEQ ID No.:l) and PMH-18 (SEQ ID No.:3) and the amino acid sequences (SEQ ID No.:2 and 4, respectively) are presented in the Listings of Sequences. Proteins PMH-17 and PMH-18 can be faith. : tc & t gb & amp; able to induce the formation of cartilage, bone or other connective tissue, or combinations thereof. The cartilage and / or bone and / or other connective tissue formation activity in the rat bone formation test is described below. The proteins PMH-17 and PMH-18 can also be characterized by the ability to demonstrate effects on the growth and / or differentiation of embryonic cells and / or stem cells. In this manner, the proteins or compositions of the present invention may also be useful for the treatment of cell populations, such as populations of embryonic cells or pluripotent cells, to enhance or enrich the growth and / or differentiation of cells. Alternatively, the proteins or compositions of the present invention may also be useful for the maintenance of a population of cells including populations of differentiated cells, for example, neuronal cells, epithelial cells, dendritic cells, chondrocytes, osteocytes, muscle cells or other cells. differentiated phenotypes. Human PMH-17 proteins, can be produced by culturing a transformed cell with a DNA sequence containing nucleotides, a DNA sequence encoding the mature PMH-17 polypeptide, containing nucleotide # 427 to nucleotide # 1098 as shown in SEQ ID No. : ly and recovering and purifying, from the culture medium, a protein characterized by the amino acid sequence containing amino acids from # 1 to # 224 as shown in SEQ ID No.:2, substantially free of other proteinaceous materials, with which occurs jointly. For production in mammalian cells, the DNA sequence further comprises a DNA sequence encoding a 5 'propeptide suitable for and linked in frame to the nucleotide sequence encoding the mature PMH-17 related polypeptide. Human PMH-18 proteins can be produced by culturing a transformed cell with a DNA sequence containing nucleotides, a DNA sequence encoding the mature PMH-18 polypeptide, containing nucleotide # 406 to nucleotide # 1098 as shown in FIG. shows in SEQ ID No .: 3 and recovering and purifying, from the culture medium, a protein characterized by the amino acid sequence containing amino acids from # 1 to # 231, as shown in SEQ ID No .: 4 , substantially free of other proteinaceous materials with which it is produced together. For production in mammalian cells, the DNA sequence further contains a DNA sequence encoding a 5 'propeptide suitable for and linked in frame to the nucleotide sequence encoding the mature PMH-18 related polypeptide. The propeptide may be the propeptide related to native PMH-17 or PMH-18, or it may be a propeptide from another protein of the TGF-β superfamily. Where the native PMH-17 and PMH-18 propeptide is used, human PMH-17 and PMH-18 can be produced by culturing a transformed cell with a DNA sequence containing a DNA sequence encoding the PMH polypeptide. -17 and complete PMH-18, which contain nucleotides from # 1 to # 1098 as shown in SEQ ID No.:l, or nucleotides from # 1 to # 1098 as shown in SEQ ID No. : 3, producing a protein characterized by the amino acid sequence that contains the amino acids from # -142 to # 224 as shown in SEQ ID No.:2, of which, the amino acids from # -142 to # -1 contain the native propeptide of human PMH-17; or # - 5 135 to # 231 of SEQ ID No.:4 of which the amino acids of # -135 to # -1 contain the propeptide native to human PMH-18 and recovering and purifying from the culture medium, a protein characterized by the amino acid sequence containing amino acids from # 1 to # 224, as shown in SEQ ID No.:2, or from # 1 to # 231 of SEQ ID No.:4, respectively, substantially free of other proteinaceous materials with which it is produced together. Based, in part, on the von Heginje signal peptide prediction algorithm, approximately first 17 to 23 amino acids of SEQ ID No.:2 and 4, appear to be involved in signaling the secretion of the mature peptide. Accordingly, in one embodiment of the invention, the DNA encoding a signal peptide, such as the signal peptide of native PMH-17 or PMH-18, or another peptide The recognized signal can be linked directly to the sequence encoding the peptide PMH-17 or mature PMH-18. It is expected that other species, particularly the human one, have DNA sequences homologous to the human PMH-17 and PMH-18 proteins. Therefore, the invention includes Methods for obtaining the DNA sequences encoding the human PMH-17 and PMH-18 proteins, the DNA sequences obtained by those methods, and the human proteins encoded by those DNA sequences. This method involves using the nucleotide sequences of PMH-17 and PMH-18 human, or portions thereof, to design probes for . - ** ¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡Selective screening of libraries for the corresponding gene of other species or 'Sequences encoding or fragments of the same, from the use of standard techniques. In this manner, the present invention can include DNA sequences from other species that are homologous to human PMH-17 and PMH-18 proteins, and can be obtained using the human PMH-17 and / or PMH-18 sequences. The present invention may also include functional fragments of the human PMH-17 and PMH-18 proteins and DNA sequences encoding those functional fragments, as well as functional fragments of other related proteins. The ability of that fragment, to function, can be determined by assaying the protein in biological assays described for the assay of proteins PMH-17 and PMH-18. Disclosed herein are DNA sequences encoding the PMH-17 protein (SEQ ID No.:l) and the human PMH-18 protein (SEQ ID No .: 3), mature, complete, and the corresponding amino acid sequences (SEQ ID No.:2 and 4, respectively). The PMH-17 and PMH-18 proteins of the present invention, such as human PMH-17 and PMH-18, can be produced by culturing a cell transformed with the correlator DNA sequence, such as the DNA sequence of Human PMH-17 and PMH-18, and the recovery and purification of the protein, such as PMH-17 or PMH-18, from the culture medium. The purified protein, expressed, is substantially free of other proteinaceous materials with which it is produced together, as well as other contaminants. It is contemplated that the purified, recovered protein exhibits cartilage and / or bone or connective tissue formation activity. In this way, the proteins of the invention can be further characterized by the ability to demonstrate cartilage and / or bone and / or other connective tissue activity in the bone formation test in rat, described below. The proteins PMH-17 and PMH-18 can be further characterized by the ability to demonstrate effects on the growth and / or differentiation of embryonic cells and / or stem cells. In this way, the proteins or compositions of the present invention can be further characterized by their ability to improve or enrich the growth and / or differentiation of the cells. Another aspect of the invention provides pharmaceutical compositions containing a therapeutically effective amount of PMH-17 and / or human PMH-18 protein, in a pharmaceutically acceptable carrier or vehicle. These compositions of the invention can be used in bone formation. These compositions can also be used for the formation of cartilage, or other connective tissue, including tendon, ligament, meniscus and other connective tissue, as well as combinations of the above, for example the regeneration of the tendon-to-bone fixation apparatus. The compositions of the present invention, such as the compositions of PMH-17 and / or human PMH-18, can also be used for wound healing and tissue repair. The compositions of the invention may further include at least one other therapeutically useful agent, such as the proteins PMH, PMH-1, PMH-2, PMH-3, PMH-4, PMH-5, PMH-β and PMH-7, described for example in U.S. Patents 5,108,922; 5,013,649, 5,116,738; 5,106,748; 5,187,076; and 5,141,905; The PMH-8 described in PCT publication O91 / 18098; and PMH-9, described in PCT publication O93 / 00432, PMH-10, described in U.S. Patent 5,637,480; PMH-11, described in U.S. Patent 5,639,638 or PMH-12 or PMH-13 described in U.S. Patent 5,658,882, PMH-15, described in U.S. Patent 5,635,372 and PMH-16 described in the co-pending patent application serial number 08 / 715,202. Other compositions that may also be useful include Vgr-2, and any of the growth and differentiation factors [GDF], including those described in PCT applications W094 / 15965; W094 / 15949; WO95 / 01801; O95 / 01802; W094 / 21681; 094/15966; WO95 / 10539; WO96 / 01845; O96 / 02559 and others. Also useful in the present invention may be the BIP, described in WO94 / 01557; HPP00269, described in Japanese publication number: 7-250688; and MP52, described in PCT application O93 / 16099. Descriptions of all applications are incorporated herein by reference. The compositions of the invention may comprise, in addition to a protein related to PMH-17 and / or PMH-18, other therapeutically useful agents that include growth factors such as epidermal growth factor (EGF), growth factor of fibroblasts (FGF), transforming growth factor (TGF-a and TGF-β), activins, inhibins and insulin-like growth factor (IGF). The compositions may also include, for example, an appropriate matrix, for the support of the composition and provide a surface for the growth of bone and / or cartilage and / or - ^ 5 ^ j ^^ fe ^ &fc ^ j¿Siíli? = Aij other connective tissue. The matrix can provide a slow release of the osteoinductive protein and / or the appropriate environment for the presentation thereof. The compositions containing PMH-17 and / or PMH-18, can be used in methods for the treatment of a number of defects in bone and / or cartilage and / or other connective tissue, for the treatment of periodontal disease and for the healing of various types of tissues and wounds. The tissue and wounds that can be treated include the epidermis, nerve, muscle, including the heart muscle and other tissues and wounds, or tissue from other organs such as the liver, lung, epithelium, brain, spleen, heart, pancreas and kidney. . These methods according to the invention entail administering to a patient in need of such formation of bone and / or cartilage and / or other connective tissue, wound healing or tissue repair, an effective amount of a PMH- protein. 17 and / or PMH-18. Compositions containing PMH-17 and / or PMH-18 can also be used for the treatment or prevention of conditions such as osteoarthritis, osteoporosis, and other abnormalities of bone, cartilage, muscle, tendon, ligament, or other tissue. connective, or tissue of other organs such as the liver, lung, epithelium, brain, spleen, heart, pancreas and kidney. These methods can also lead to the administration of a protein of the invention, together with at least one other PMH protein, as described above. In addition, these methods may also include the administration of a PMH-17 and / or PMH-18 protein with other growth factors including EGF, FGF, TGF-α, TGF-β, activin, inhibin and IGF. (f ^^ W ^? ^ '^ ^ i ^ W ^ I ^ 1 Still a further aspect of the invention are the DNA sequences encoding the expression of a protein PMH-17 and / or PMH-18. include the nucleotide sequence in a 5 'to 3' direction illustrated in SEQ ID NO: SEQ ID NO: 3, DNA sequences which, but for the degeneracy of the genetic code, are identical to the sequence of DNA, SEQ ID No.: SEQ ID No.:3, and encode the protein of SEQ ID No .: 2 or SEQ ID No.:4. Also included in the present invention are the DNA sequences that are hybridized under stringent conditions with the DNA sequence of SEQ ID NO: SEQ ID NO: 3 and encode a protein that has the ability to induce the formation of cartilage, bone and / or other connective tissue, or tissue of other organs such as the liver, lung, epithelium, brain, spleen, heart, pancreas and kidney, or other activities described for PMH-17 and PMH-18. efferides include those that hybridize under stringent conditions [eg, see the conditions described in Maniatis et al, Molecular Cloning (A Laboratory Manual), Cold Spring Harbor Laboratory (1982), pages 387 to 389], It is generally preferred that those sequences of DNA encode a polypeptide that is at least about 80% homologous, and more preferably, at least about 90% homologous, to the cysteine structure of the conserved C-terminus, of the amino acid sequence shown in SEQ ID No. : 2 or SEQ ID No.:4. Finally, the allelic variations or other variations, of the sequences of SEQ ID No. 1 or SEQ ID No. 3, whether those nucleotide changes result or not, changes ? »^ Z * H & i¿ect3ate? i ~ > . in the peptide sequence, wherein the peptide sequence retains one or more of the activities of PMH-17 and / or PMH-18, are also included in the present invention. The present invention also includes fragments of the DNA sequence of PMH-17 and / or PMH-18 shown in SEQ ID NO: SEQ ID NO: 3 which encodes a polypeptide that preserves the activity of the PMH-protein. 17 and / or PMH-18. The DNA sequences of the present invention are useful, for example, as probes for the detection of mRNA encoding PMH-17 and / or PMH-18 in a given population of cells. In this way, the present invention includes methods of detecting or diagnosing genetic disorders involving the genes of PMH-17 and / or PMH-18, or disorders involving cellular, organ, or tissue disorders, in which PMH-17 and / or or PMH-18 is transcribed or expressed irregularly. The DNA sequences may also be useful for the preparation of vectors for applications in gene therapy as described below. A further aspect of the invention includes vectors containing a DNA sequence as described above, in functional association with an expression control sequence for the same. These vectors can be used in a novel process to produce a PMH-17 and / or PMH-18 protein of the invention, in which a cell line transformed with a DNA sequence encoding a PMH-17 and / or PMH-protein 18, in functional association with an expression control sequence for the same, is grown in a suitable culture medium and a protein related to PMH-17 and / or PMH-18, is recovered and purified thereof. This process can employ a number of known cells, both prokaryotic and eukaryotic, and host cells, for the expression of the polypeptide. The vectors can be used in gene therapy applications. In that use, the vectors can be transfected into cells of an ex vivo patient, and the cells can be reintroduced to a patient. Alternatively, the vectors can be introduced to a patient in vivo through target-directed transfection. Still a further aspect of the invention are the proteins or polypeptides PMH-17 and / or PMH-18. These polypeptides are characterized by having an amino acid sequence that includes the sequence illustrated in SEQ ID No.:2 or SEQ ID No .: 4, variants of the amino acid sequence of SEQ ID No.:2 or SEQ ID No. .: 4, including natural allelic variants, and other variants in which the protein retains the ability to induce the formation of cartilage and / or bone and / or other connective tissue, or tissue from other organs such as the liver, lung , epithelium, brain, spleen, heart, pancreas and kidney, or other activity characteristic of PMH-17 and / or PMH-18. Preferred polypeptides include a polypeptide that is at least about 80% homologous, and more preferably at least about 90% homologous, to the amino acid sequence of mature, human PMH-17 and / or PMH-18, shown in SEQ ID. No .: 2 or SEQ ID No .: 4. Finally, the allelic variations or other variations of the sequences of SEQ ID No.:2 or SEQ ID No.:4, whether these amino acid changes are induced by mutagenesis , chemical alteration, or alteration of the DNA sequence used to produce the polypeptide, wherein the peptide sequence S? Sdtei ** ¿2¿ »A ^ = * '* - "still have activity of PMH-17 and / or PMH-18, are also included in the present invention. The present invention also includes fragments of the amino acid sequence of PMH-17 and / or PMH-18, shown in SEQ ID No.:2 or SEQ ID No .: 4, which preserve the activity of the PMH-17 protein and / or PMH-18. The purified proteins of the present invention can be used to generate antibodies, either monoclonal or polyclonal, for PMH-17 and / or PMH-18.

Human, and / or other proteins related to PMH-17 or PMH-18, using methods that are known in the art of antibody production. In this manner, the present invention also includes antibodies to human PMH-17 and / or PMH-18, and / or other related proteins. Antibodies may be useful for the purification of PMH-17 and / or PMH-18 and / or other proteins related to PMH-17 and / or PMH-18, or to inhibit or prevent the effects of proteins related to PMH -17 and / or PMH-18. Protein PMH-17 and / or PMH-18, and related proteins, may be useful for induce the growth and / or differentiation of embryonic cells and / or pluripotent cells. In this way, the proteins or compositions of the present invention may also be useful for the treatment of relatively undifferentiated cell populations, such as populations of embryonic cells or pluripotent cells, to enhance or enrich the growth and / or differentiation of cells. The populations of treated cells can be useful for implantation and for applications in gene therapy. 30 Description of the Sequences SEQ ID No. : 1 is a nucleotide sequence containing nucleotide sequences encoding the entire mature human PMH-17 polypeptide. SEQ ID No .: 2 is an amino acid sequence that contains the mature human PMH-17 polypeptide sequence. SEQ ID No .: 3 is a nucleotide sequence that contains a nucleotide sequence that encodes all mature human PMH-18 polypeptide. SEQ ID No .: 4 is an amino acid sequence that contains the mature human PMH-17 polypeptide sequence.

DETAILED DESCRIPTION OF THE INVENTION The human PMH-17 and PMH-18 sequences of the present invention can be obtained using the entire murine Lefty DNA sequence, or fragments thereof or a partial sequence of human PMH-17 or PMH-18 , as a probe. In this manner, the DNA sequence of human PMH-17 and PMH-18, comprises the DNA sequence of nucleotides from # 1 to # 1098 of SEQ ID No.:lo from # 1 to # 1098 of SEQ ID No.:3 The human PMH-17 and PMH-18 proteins contain the amino acid sequences of # -142 to # 224 of SEQ ID No.:2, or # -135 to # 231 of SEQ ID No .: 4, respectively . The mature human PMH-17 and PMH-18 proteins are encoded by nucleotides of # 427 to # 1098 of SEQ ID No.:l and # 406 and # 1098 of the .- < »R SEQ ID NO: 3, respectively, and contain the amino acid sequence of # 1 to # 224 of SEQ ID No.:2, or # 1 to # 231 of SEQ ID No.:4, respectively. It is to be expected that the human PMH-17 and PMH-18 polypeptides, as expressed in mammalian cells such as CHO cells, exist as a heterogeneous population of active species of PMH-17 and PMH-18 proteins with variable N terms. It is expected that the active species will comprise an amino acid sequence beginning with the residue at amino acid # 109 or # 124 of SEQ ID No.:2 or # 116 or # 131 of SEQ ID No .: 4, respectively, or it will comprise an additional sequence of amino acids, furthermore, in the direction of the N-terminus. In this manner, it is expected that the DNA sequences encoding the active PMH-17 and PMH-18 polypeptides will comprise a nucleotide sequence comprising the nucleotides # 1, # 232, # 406, # 427, # 751 or # 796 to # 1059 or # 1098 of SEQ ID No.:l, or # 1, # 232, # 406, # 427, # 751 or # 796 to # 1059 or # 1098 of SEQ ID No .: 3, respectively. Therefore, it is expected that the active species of human PMH-17 and PMH-18, include those that comprise amino acids # -142, # -65, # -7, # 1, # 109 or # 124 to # 211 or # 224 of SEQ ID No.:2, or # -135, # -58, # 1, # 8, # 116 or # 131 to # 218 or # 231 of SEQ ID No.:4, respectively. A host cell can be transformed with a coding sequence that encodes a propeptide suitable for the secretion of proteins by the host cell, which is linked in the appropriate reading frame, to the coding sequence for the protein PMH-17 and PMH- 18 mature. For example, see U.S. Patent No. 5,658,882, in which the PMH-2 propeptide is fused to DNA encoding a mature PMH-12 protein. The description of this reference is incorporated herein by reference. In this manner, the present invention includes chimeric DNA molecules, which contain a DNA sequence encoding a propeptide from a member of the TGF-β superfamily of proteins, other than PMH-17 and PMH-18, which is linked to a correct reading frame, to a DNA sequence encoding the human PMH-17 or PMH-18 protein, or a related protein. The term "chimeric" is used to mean that the propeptide originates from a different polypeptide than the native PMH-17 or PMH-18 protein. The N-terminus of an active species of human PMH-17 is expected to be determined experimentally by expression in E. coli as follows: [M] ARVTV. Thus it seems that the N-terminus of this species of PMH-17 is in amino acid # 1 of SEQ ID No.:l, and a DNA sequence encoding that species of PMH-17, comprising the nucleotides of # 427 to # 1098 of SEQ ID No.:l. The apparent molecular weight of the human PMH-17 monomer is expected to be determined experimentally by SDS-PAGE, as approximately 24.8 kD in a 16% tricine gel, from Novex. It is expected that the human PMH-17 protein exists as a clear and colorless solution, in 0.1% trifluoroacetic acid. It is expected that other PMH-17 proteins, as expressed by mammalian cells such as CHO cells, also exist as a heterogeneous population. j AmjL? aA ¿? > * * * Of active species of the protein related to PMH-17, with variable N terms. For example, it is expected that the active species of the human PMH-17 protein will comprise an amino acid sequence beginning with the residue 5 cysteine at amino acid # 109 or the glutamic acid residue at position # 124 of SEQ ID No .: 2, or will further comprise an additional amino acid sequence in the direction of the N-terminus. Thus, DNA sequences encoding the PMH-17 proteins are expected.

Active, include those comprising a nucleotide sequence containing nucleotides # 1, 232, 406, 427, 751 or 796 to # 1059 or # 1098 of SEQ ID No.:l. Accordingly, active human PMH-17 proteins include those comprising amino acids # -142, -65, -7, 1, 109 or 124 to # 211 or 224 of SEQ ID No.:2. The N-terminus of an active species of human PMH-18 is expected to be determined experimentally by the expression in E. coli as follows: [M] LSPRS. In this way, it seems that the N term of this species of PMH-18, is in the amino acid # 1 of SEQ ID No .: 3, and a DNA sequence encoding that PMH-18 species, comprising nucleotides of # 406 to # 1098 of SEQ ID NO: 3. The apparent molecular weight of the PMH-18 monomer is expected to be experimentally determined by SDS-PAGE as approximately 25.6 kD in a 16% tricine gel, from Novex. It is expected that the human PMH-18 protein, exists as a clear and colorless solution, in 0.1% trifluoroacetic acid. Other PMH-18 proteins are expected, such as are expressed by mammalian cells such as CHO cells, also exist as a heterogeneous population i "* S; * '4 ± SS =, -:» & ^ üíi. of active protein species related to PMH-18, with variable N terms. For example, it is expected that the active species of human PMH-PMH-18 protein will comprise an amino acid sequence beginning with the 5-cysteine residue at amino acid # 116 or with the glutamic acid residue at position # 131 of the SEQ ID NO: 4, or will further comprise an additional amino acid sequence in the N-terminal direction. Thus, it is expected that the DNA sequences encoding the active PMH-18 0 proteins will include those containing a sequence. of nucleotides containing nucleotides # 1, 232, 406, 427, 751 or 796 to # 1059 or # 1098 of SEQ ID No.:3. Accordingly, active, human PMH-18 proteins include those which contain the amino acids of # -135, -58, 1, 8, 5 116 or 131 to # 218 or 231 of SEQ ID No.:4. The PMH-17 or PMH-18 proteins of the present invention, include polypeptides having a molecular weight of about 24.8 to 25.6 kD in monomeric form, and the polypeptide contains the amino acid sequence of SEQ ID 0 No.:2 or SEQ ID No.:4, respectively, and has the ability to induce the formation of cartilage and / or bone or other connective tissue, in the Sampath-Reddi ectopic implant assay modified by Rosen, described in the examples. The PMH-17 or PMH-18 proteins recovered from the culture medium are purified by isolating them from other proteinaceous materials with which they are produced together, and from other contaminants present. The proteins PMH-17 or PMH-18 can be characterized by the ability to induce the formation of cartilage and / or bone 0 and / or other connective tissue, and by the repair and differentiation of • "- gf" "'^ M ^ m ^ w ^ mum s i1 .. another type of tissue, for example, in the rat bone formation test, described later. In addition, the PMH-17 or PMH-18 proteins can be further characterized by their effects on the growth and / or differentiation of embryonic cells and / or stem cells. In this way, the proteins or compositions of the present invention can be characterized by the assay of embryonic, pluripotent cells, described later. The PMH-17 or PMH-18 proteins provided herein also include factors encoded by sequences similar to those of SEQ ID NO: SEQ ID NO: 3, but in which modifications or deletions are naturally provided (for example, example, allelic variations in the nucleotide sequence that can result in changes in the amino acids in the polypeptide) or deliberately manipulated by engineering. For example, synthetic polypeptides can duplicate, in whole or in part, continuous sequences of the amino acid residues of SEQ ID No .: 2 or SEQ ID No.:4. These sequences, by virtue of sharing structural and conformational characteristics, primary, secondary or tertiary, with the bone growth factor polypeptides of SEQ ID No .: 2 or SEQ ID No .:, may possess biological properties in common with them. It is known, for example, that numerous conservative amino acid substitutions are possible, without significantly modifying the structure and conformation of a protein, thus maintaining the biological properties as such. For example, it is recognized that conservative amino acid substitutions can be made between amino acids with basic side chains, such as lysine (Lys or K), arginine (Arg or R) and histidine (His or H); amino acids with acid side chains, such as aspartic acid (Asp or D) and glutamic acid (Glu or E); 5 amino acids with polar, uncharged side chains, such as asparagine (Asn or N), glutamine (Gln or Q), serine (Ser or S), threonine (Thr or T), and tyrosine (Tyr or Y) and amino acids with non-polar side chains, such as alanine (Ala or A), glycine (Gly or G), valine (Val or V), leucine (Leu or L), isoleucine (lie or í), proline (Pro or P), phenylalanine (Phe or F), methionine (Met or M), tryptophan (Trp o) and cysteine (Cys or C). In this way, these modifications and deletions of the native PMH-17 or PMH-18 can be used as substituents biologically active for PMH-17 or natural PMH-18 and other polypeptides, in therapeutic processes. It can be easily determined if a certain variant of PMH-17 or PMH-18 maintains the biological activity of PMH-17 or PMH-18, subjecting PMH-17 or PMH-18 and the variant of PMH-17 or PMH-20 18 to the assays described in the examples. Other specific mutations of the sequences of the PMH-17 or PMH-18 proteins described herein, involve modifications of the glycosylation sites. These modifications may involve sites of O-linked or N-linked glycosylation. For example, the absence of glycosylation, or only partial glycosylation, results in the substitution or deletion of amino acids at asparagin-linked glycosylation recognition sites. The recognition sites of glycosylation linked to asparagine comprise sequences ^ ^ f ^ '? e ^^' ^ s0? ^?. of tripeptides that are specifically recognized by the appropriate cell glycosylation enzymes. These tripeptide sequences are, or asparagine-X-threonine or asparagine-X-serine where X is usually any amino acid. A variety of substitutions or deletions of amino acids in one or both of the first or third amino acid positions of a glycosylation recognition site (and / or deletion of amino acids in the second position) results in non-glycosylation in the sequence of modified tripeptides. Additionally, bacterial expression of the protein related to PMH-17 and PMH-18 will also result in the production of a non-glycosylated protein, even if the glycosylation sites are left unmodified. The present invention also encompasses the novel DNA sequences, without association with DNA sequences encoding other proteinaceous materials and encoding for the expression of the PMH-17 and PMH-18 proteins. These DNA sequences include those depicted in SEQ ID. No.: SEQ ID No .: 3 in a 5 'to 3' direction and those sequences that hybridize thereto under stringent washing conditions for hybridization [eg, 0. IX SSC, 0.1% SDS at 65 ° C; see, T. Maniatis et al, Molecular Cloning (A Laboratory Manual), Cold Spring Harbor Laboratory (1982), pages 387 to 389] and encode a protein that maintains one or more of the activities described herein for the PMH-? 7 or PMH-18. These DNA sequences also include those that comprise the DNA sequence of SEQ ID No.:l or SEQ ID No.:3 and those that hybridize thereto under conditions of astringent hybridization and that encode a protein that maintains the other activities described for PMH-17 or PMH-18. Similarly, the DNA sequences encoding the PMH-17 or PMH-18 polypeptides encoded by the sequences of SEQ ID No.:SE SEQ ID No.:3, respectively, or the PMH-17 or PMH- polypeptides 18 which comprise the amino acid sequence of SEQ ID No .: 2 or SEQ ID No .: 4, but which differ in the sequence of codons due to degenerations of the genetic code or due to allelic variations (changes of bases, natural , in the population of species, which may or may not result in a change in amino acids) also encode the novel factors described herein. Variations in the DNA sequences of SEQ ID No.: SEQ ID No.:3 that are caused by point mutations or by induced modifications (including insertion, deletion and substitution) to enhance activity, life medium or the production of the encoded polypeptides are also included in the invention. Another aspect of the present invention provides a novel method for producing PMH-17 or PMH-18 polypeptides. The method of the present invention involves cultivating a suitable cell line, which has been transformed with a DNA sequence encoding a PMH-17 or PMH-18 polypeptide of the invention, under the control of known regulatory sequences. The transformed host cells are cultured and the polypeptides PMH-17 and PMH-18 are recovered and purified from the culture medium. The purified polypeptides are substantially free of other proteins with which they were produced together, as well as other contaminants. Suitable cells or cell lines can be mammalian cells, such as Chinese Hamster Ovary (CHO) cells. The selection of suitable mammalian host cells, and methods for transformation, culture, amplification, selective screening, production and purification of the product, are known in the art. See, for example, Gething and Sambrook, Nature, 293: 620-625 (1981), or alternatively, Kaufman et al, Mol. Cell. Biol., 5 (7): 1650 (1985) or Howley et al, U.S. Patent 4,419,446. Another suitable mammalian cell line, which is described in the appended examples, is the COS-1 monkey cell line. The mammalian CV-1 cell may also be suitable. Bacterial cells can also be suitable hosts. For example, different strains of E. coli (eg, HB101, MC1061) are well known as host cells in the field of biotechnology. Various strains of B. subtilis, Pseudomonas, other bacilli and the like can also be used in this method. For the expression of the protein in bacterial cells, the DNA encoding the propeptide of PMH-17 or PMH-18 is not generally necessary. Many strains of yeast cells known to those skilled in the art can also be found available as host cells for the expression of the polypeptides of the present invention. Additionally, where desired, insect cells can be used as host cells, in the method of the present invention. See, for example, Miller et al, Genetic Engineering, 8: 277-298 ff '«rU» l', "" lllti > and r "~ 'Wr? ryij? ra -ijl ^ á &ampi? fei-, (Plenum Press 1986) and the references cited therein. Another aspect of the present invention provides vectors for use in the method of expression of these novel polypeptides PMH-17 or PMH-18. Preferably the vectors contain the novel complete DNA sequences, described above, which encode the novel factors of the invention. Additionally, the vectors contain appropriate expression control sequences, allowing the expression of the PMH-17 and PMH-18 polypeptide sequences. Alternatively, vectors incorporating modified sequences, such as those described above, are also embodiments of the present invention. Additionally, the sequence of SEQ ID No.:SE SEQ ID No .: 3 or other sequences encoding the PMH-17 or PMH-18 polypeptides could be manipulated to express a mature PMH-17 or PMH-18 polypeptide, enating the Progestin PMH-17 or PMH-18 sequences and replacing them with sequences that encode the complete propeptides of other PMH proteins or members of the TGF-β superfamily. In this manner the present invention includes chimeric DNA molecules encoding a propeptide of a member of the TGF-β superfamily linked in a correct reading frame to a DNA sequence encoding a PMH-17 or PMH-18 polypeptide. The vectors can be used in the cell line transformation method and contain selected regulatory sequences, in functional association with the DNA coding sequences of the invention, which are capable of directing the replication and expression thereof in the selected host cells. The regulatory sequences for these vectors are understood by those skilled in the art "and can be selected depending on the host cells.That selection is routine and does not form part of the present invention.A protein of the present invention, which induces the formation of cartilage and / or bone and / or other connective tissue, in circumstances where that tissue does not normally appear, has application in the healing of fractures of bones and defects in cartilage or other connective tissues, in humans and in others This preparation using a PMH-17 or PMH-18 polypeptide can have prophylactic use in the reduction of closed fractures as well as in open fractures, and also in the improved fixation of artificial joints.New bone formation induced by an agent osteogenic, contributes to the repair of congenital craniofacial defects, induced by traumas, or induced by oncological resection, and is also Useful in cosmetic plastic surgery. A polypeptide related to PMH-17 or PMH-18 may be used in the treatment of periodontal disease and in other processes of tooth repair. These agents can provide an environment to attract bone-forming cells, stimulate the growth of bone-forming cells or induce differentiation of bone-forming cell progenitors, and can also support the regeneration of the periodontal ligament and bone structure. fixation device, which connects the bone and the teeth. The PMH-17 or PMH-18 polypeptides of the invention may also be useful in the treatment of osteoporosis. It has been ^^ - * ^^^ described a variety of osteogenic factors, cartilage inducers and bone inducers. See, for example, European Patent Applications 148,155 and 169,016 for analysis thereof. The proteins of the invention can also be used in the healing of wounds and in the repair of related tissue. Wound types include, but are not ted to, burns, incisions and ulcers. (See, for example, PCT publication WO84 / 01106 for the analysis of wound healing and repair of related tissue). It is further contemplated that the proteins of the invention may increase the survival of astrocytic and equal neuronal cells and therefore may be useful in the transplantation and treatment of conditions that exhibit a decrease in neuronal survival and repair. The proteins of the invention may also be useful for the treatment of conditions related to other types of tissue, such as nerve, epidermis, muscle, and tissue from other organs such as the liver, lung, epithelium, brain, spleen, heart, pancreas, and kidney. The proteins of the present invention may also be useful for the treatment of relatively undifferentiated cell populations, such as embryonic cells, or stem cells, to enhance the growth and / or differentiation of cells. The proteins of the present invention may also have value as a dietary supplement or as a component of cell culture media. For this use, the proteins can be used intact, or can be pre-digested to provide a supplement that is more easily absorbed. ^ ** \, fto ~ f The proteins of the invention may also have other useful properties, characteristic of the TGF-β superfamily of proteins. These properties include angiogenic, chemotactic and / or chemoattractant properties and effects on cells, including the induction of collagen synthesis, fibrosis, differentiation responses, cellular proliferative responses and responses that involve cell adhesion, migration and extracellular matrices. These properties make the proteins of the invention potential agents for wound healing, for the reduction of fibrosis and for the reduction of scar tissue formation. When dimerized as a homodimer or as a heterodimer with other PMH, with other members of the TGF-β superfamily of proteins, or with inhibin-a proteins or inhibin-β proteins, the heterodimer PMH-17 or PMH-18 is expected , present effects on the production of follicle stimulating hormone (FSH), as further described herein. It is recognized that SFH stimulates the development of ova in mammals ovaries (Ross et al, in Textbook of Endocrinology, ed Williams, page 355 (1981) and that excessive ovarian stimulation with FSH will lead to multiple ovulations FSH is also important in testicular function, so PMH-17 or PMH-18 can be useful as a contraceptive based on the ability of inhibins, to decrease fertility in female mammals and to decrease spermatogenesis In male mammals, administration of sufficient amounts of other inhibins may induce infertility in mammals.PHM-17 or PMH-18 may also be useful as a fertility-inducing therapeutic agent, based on the ability of activin molecules to stimulate the release of FSH from the anterior pituitary cells, see, for example, U.S. Patent No. 4,798,885, PMH-17 or PMH-18 may also be useful for the advancement of beginning of fertility in sexually immature mammals, in order to increase reproductive performance in life, of domestic animals such as cows, sheep and pigs. It is further contemplated that PMH-17 or PMH-18 may be useful in the modulation of hematopoiesis, inducing the differentiation of erythroid cells [see, for example, Broxmeyer et al, Proc. Na ti. Acad. Sci. United States of America, 85: 9052-9056 (1988) or Eto et al, Biochem. Biophys. Res. Comm. , 142: 1095-1103 (1987)], to inhibit the development of gonadal tumors [see, for example, Matsuk et al., Na ture, 360: 313-319 (1992)], or to increase the activity of the proteins morphogenetic of the bones [see, for example, Ogawa et al., J. Biol. Chem. , 257: 14233-14237 (1992)]. The PMH-17 and PMH-18 proteins may also be characterized by their ability to modulate the release of follicle stimulating hormone (FSH) in established, in vitro biological assays using rat anterior pituitary cells, as described in FIG. describes [see, for example, Vale et al, Endocrinology, 9 ^: 562-572 (1972); Ling et al., Na ture, 321: 779-782 (1986) or Vale et al., Na ture, 321: 776-779 (1986)]. It is contemplated that the PMH-17 or PMH-18 protein of the invention, when composed as a heterodimer with inhibin α or inhibin β chains, will exhibit regulatory effects, either stimulatory or inhibitory, on the release of the follicle stimulating hormone ( FSH), from anterior pituitary cells as described [Ling et al, Nature, 321: 779-782 (1986) or Vale et al., Na ture, 321: 776-779 (1986); Vale et al, Endocrinology, 91: 562-572 (1972)]. Therefore, depending on the particular composition, it is expected that the PMH-17 or PMH-18 protein of the invention may have contrasting and opposite effects in the release of follicle stimulating hormone (FSH) from the anterior pituitary. It has been shown that activin A (the homodimeric composition of inhibin βA) has erythropoietic-stimulant activity [see for example, Eto et al, Biochem. Biophys. Res. Commun. , 142: 1095-1103 (1987) and Murata et al, Proc. Na ti. Acad. Sci. United States of America, 85: 2434-2438 (1988) and Yu et al, Na ture, 330: 765-767 (1987)]. It is contemplated that the PMH-17 and PMH-18 proteins of the invention may have a similar erythropoietic-stimulatory activity. This activity of the proteins PMH-17 and PMH-18 can be further characterized by the ability of the proteins PMH-17 and PMH-18 to demonstrate erythropoietin activity, in the biological assay performed using the human K-562 cell line, as described [Lozzio et al., Blood, 4_5: 321-334 (1975) and U.S. Patent No. 5, 071, 834]. A further aspect of the invention is a method and therapeutic composition, for repairing fractures and other conditions related to defects in cartilage and / or bone and / or other connective tissue, or periodontal diseases. The invention further comprises methods and therapeutic compositions for wound healing and tissue repair. These compositions comprise a therapeutically effective activity of at least one of the proteins related to PMH-17 or PMH-18, of the invention, in admixture with a pharmaceutically acceptable carrier, carrier or matrix. It is further contemplated that the compositions of the invention may increase neuronal survival and therefore be useful in the transplantation and treatment of conditions exhibiting a decrease in neuronal survival. The compositions of the invention may further include at least one other therapeutically useful agent, such as members of the protein PGF-β superfamily, which includes the proteins PMH, PMH-1, PMH-2, PMH-3, PMH- 4, PMH-5, PMH-6, PMH-7, described for example in U.S. Patents 5,108,922; 5,013,649; 5,116,738; 5,106,748; 5,187,076; and 5,141,905; PMH-8 described in PCT publication WO91 / 18098; and PMH-9, described in U.S. Patent 5,661,007; PMH-10, described in U.S. Patent 5,637,480; PMH-11, described in U.S. Patent 5,639,638 or PMH-12 or PMH-13 described in U.S. Patent 5,658,882; or PMH-15, described in U.S. Patent 5,635,372; or the PMH-16 described in the copending patent application Serial No. 08 / 715,202. Other compositions that may also be useful include Vgr-2, and any of the growth and differentiation factors [GDF], which include those described in PCT applications W094 / 15965, W094 / 15949; WO95 / 01801; WO95 / 01802; W094 / 21681; W094 / 15966; WO95 / 10539; WO96 / 01845; ? *.: ± WO96 / 02559 and others. Also useful in the present invention may be the BIP, described in WO94 / 15057; HPP00269, described in Japanese publication number: 7-250688; and MP52, described in PCT application WO93 / 16099. Descriptions of all applications are incorporated herein by reference. It is expected that the human PMH-17 and PMH-18 protein may exist in nature as monomers, or as homodimers or heterodimers. In order to promote the formation of dimers of PMH-17 or PMH-18 and of useful proteins with increased stability, the DNA sequence of SEQ ID NO: SEQ ID No .: 3 can be engineered to provide one or more additional cysteine residues to increase the formation of the potential dimer. The resulting DNA sequence would be capable of producing an "added cysteine variant" of PMH-17 or PMH-18. In a preferred embodiment, the DNA sequence of SEQ ID No.: SEQ ID No. 3 is engineered, so that one or more codons can be altered in a nucleotide triplet, encoding a residue of cysteine, such as TGT or TGC. Alternatively, "added cysteine variants" of the PMH-17 or PMH-18 protein can be produced by altering the protein sequence at the amino acid level by altering one or more of the amino acid residues of SEQ ID. No.:2 or SEQ ID No.:4 to cysteine. The production of the "added cysteine variants" of proteins is described in US Pat. No. 5, 166, 322, the description of which is incorporated herein by reference. In preferred embodiments, the glutamic acid residue at position # 173 of SEQ ID No.:2 or position # 180 of SEQ ID No .: 4, is replaced by a cysteine residue. It is expected that the proteins of the invention may act in concert with, or perhaps synergistically with, other proteins and related growth factors. Therapeutic methods and additional compositions, of the invention, therefore comprise a therapeutic amount of at least one PMH-17 or PMH-18 protein of the invention, with a therapeutic amount of at least one other member of the TGF-β protein superfamily. , such as the PMH proteins described in the applications mentioned above. These combinations may comprise separate molecules of the PMH proteins or heteromolecules composed of different portions of PMH. For example, a method and composition of the invention may comprise a disulfide linked dimer comprising a subunit of PMH-17 or PMH-18 protein and a subunit of one of the "PMH" proteins described above. Thus, the present invention includes a polypeptide related to purified PMH-17 or PMH-18, which is a heterodimer wherein a subunit comprises the amino acid sequence of amino acid # 1 to amino acid # 224 of SEQ ID No. : 2 or from # 1 to # 231 of SEQ ID No .:, and a subunit comprises an amino acid sequence for a bone morphogenetic protein, selected from the group consisting of PMH-2, PMH-3, PMH- 4, PMH-5, PMH-6, PMH-7, PMH-8, PMH-9, PMH-10, PMH-11, PMH-12, PMH-13, PMH-15, or PMH-16. A further embodiment may comprise a heterodimer of portions related to PMH-17 or PMH-18, for example a heterodimer of human PMH-17 and human PMH-18, or of human PMH-17 and murine Lefty protein, which it is a homologue of human PMH-17 and PMH-18. In addition, the protein PMH-17 or PMH-18 can be combined with other beneficial agents for the treatment of the defect in bone and / or cartilage and / or other connective tissue, wound, or tissue in question. These agents include various growth factors such as epidermal growth factor (EGF), fibroblast growth factor (FGF), platelet derived growth factor (PDGF), transforming growth factors (TGF-α and TGF- ß), activmas, inhibins, fibroblast growth factor k (kFGF), parathyroid hormone (PTH), peptide related to parathyroid hormone (PTHrP), inhibitory factor of leukemia (LIB / HILA / DA), insulin growth factors (IGF-I IGF-II). Portions of these agents can also be used in compositions of the present invention. The preparation and formulation of those physiologically acceptable protein compositions, taking due account of PH, isotonicity, stability and the like, is within the skill in the art. The therapeutic compositions are also valuable for veterinary applications, due to the lack of species specificity in the PMH proteins. Particularly domestic animals and thoroughbred horses, in addition to humans, are desired patients for that treatment with the PMH-17 or PMH-18 proteins of the present invention. The therapeutic method includes administering the composition topically, systemically or locally as an implant or device. When administered, the therapeutic composition for use in this invention is, of course, in a pyrogen-free physiologically acceptable form. In addition, the composition can be encapsulated or injected, desirably, in a viscous form for administration to the site of bone, cartilage or other connective tissue or other tissue damage. Topical administration may be convenient for wound healing and tissue repair. Therapeutically useful agents, other than the PMH-17 or PMH-18 proteins, which may also optionally be included in the composition described above, may be administered, alternatively, or additionally, simultaneously or sequentially with the PMH composition, in the methods of the invention. Preferably, for the formation of bone and / or cartilage and / or other connective tissue, the composition includes a matrix capable of delivering proteins related to PMH-17 or PMH-18, or other PMH proteins, to the site of bone damage. and / or cartilage and / or other connective tissue, providing a structure for the development of bone and cartilage and other connective tissue, and which can be optimally reabsorbed in the body. The matrix can provide a PMH-17 or PMH-18 protein, or other slow-release bone-inducing protein, as well as an adequate presentation and an appropriate environment for cell infiltration. These matrices can be formed of materials that are currently in use for other implanted medical applications. The choice of matrix material is based on biocompatibility, biodegradability, mechanical properties, cosmetic appearance and interface properties. The particular application of the compositions of PMH-17 or 'j -.-, - ~ »-, jt I.IÍ].? | -. - y ^ y ^ ^ ¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡! Potential matrices for the compositions may be calcium sulfate, tricalcium phosphate, oxyapatite, polylactic acid and chemically defined and biodegradable polyanides. 5 Other potential materials are biodegradable and biologically well defined, such as bone or dermal collagen. Additional matrices are composed of pure proteins or components of the extracellular matrix. Other potential matrices are non-biodegradable and chemically defined, such as sintered oxyapatite, bioglass, aluminates, or other ceramics. The matrices can be composed of combinations of any of the types of materials mentioned above, such as polylactic acid and oxyapatite or collagen and phosphate tricalcium. The composition of bioceramics can be altered, such as in calcium aluminate-phosphate and processed to alter pore size, particle size, particle shape and biodegradability. The dosage regime will be determined by the doctor who attends the case, considering several factors that modify the action of the protein PMH-17 or PMH-18, for example, the amount of bone weight or other tissue that is desired to be formed, the site of damage to the bone or tissue, the condition of the tissue or bone damaged, the size of a wound, the type of damaged tissue, the age, sex and diet of the patient, the severity of any infection, the time of administration and other clinical factors. The dosage may vary with the type of matrix used in the reconstitution and the types of PMH proteins that are found in the composition. In general * the systemic or injectable administration will be initiated at a dose that is minimally effective, and the dose will be increased over a certain preselected time until a positive effect is observed. Subsequently, incremental increases in dosage will be made, limiting those increasing increases to such levels that produce a corresponding increase in effect, and at the same time taking into account any adverse effects that may appear. The addition of other known growth factors, such as IGF I (the insulin-like growth factor I) to the final composition, can also affect the dosage. Progress can be inspected by periodically evaluating the growth and / or repair of the bone or tissue. The progress can be inspected, for example, by X-rays, histomorphometric determinations and tetracycline labeling. The following examples illustrate the practice of the present invention with respect to the proteins human PMH-17 and human PMH-18 and other proteins related to PMH-17 and PMH-18. The skilled in the art will recognize that numerous variations and modifications are possible. These variations and modifications constitute part of the present invention.

EXAMPLES EXAMPLE 1 Isolation of DNA The DNAs encoding the human PMH-17 and PMH-18 proteins "* ^ the proteins related to human PMH-17 and PMH-18, can be isolated through several techniques known to the in the art, using the sequence information provided in the Sequence Lists, based on the knowledge of the PMH proteins and other proteins that are within the TGF-β family, the carboxyl terminal portion of these molecules is predicted. (mature peptide) exhibit greater sequence conservation than most portions of the amino terminal (propeptide region). This sequence relationship between PMH proteins and other proteins within the TGF-β family allows those skilled in the art, design DNA probes from the portion encoding the carboxyl terminus (region encoding the mature peptide) of these molecules, which can be used to identify related PMH proteins and other proteins that are within the TGF-β family. The coding region of the mature peptide, of the murine Lefty gene, can be used to identify human PMH-17 and PMH-18 proteins and related proteins. Bacteriophage AF02, which contains the DNA sequence encoding human PMH-17, has been deposited at the American Type Culture Collection, 12301 Parkla n Drive, Rockville, MD ATCC under accession # 202060, on November 24, 1997. This deposit complies with the requirements of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purpose of Procedures and "Norms thereunder." Bacteriophage AF04, which contains the DNA sequence encoding human PMH-18, has been deposited at the American Type Culture Collection, 12301 Parkla n Drive, Rockville, MD ATCC under a] S8eso # 20259, on November 24, 1997. This deposit complies with the requirements of the Budapest Treaty of the International Recognition of the Deposit of Microorganisms for the Purpose of Patent Procedures and Standards thereunder Based on knowledge of other PMH proteins and other proteins that are within the TGF-β family, it is predicted that the precursor polypeptides of human PMH-17 and PMH-18 will be cleaved in the multibasic sequence, according to a proposed consensus proteolytic processing sequence of Arg-X-X-Arg. Excision of the precursor polypeptide of human PMH-17 is expected to generate a mature peptide of 224 amino acids starting with the amino acid alanine at position # 1 of SEQ ID No .: 2 or a mature peptide of 101 amino acids beginning with the amino acid glutamic acid at position # 124 of SEQ ID No.:2. Excision of the precursor polypeptide of human PMH-18 is expected to generate a mature peptide of 231 amino acids, starting with the amino acid leucine at position # 1 of SEQ ID No .: 4, or a mature peptide of 101 amino acids , which begins with the amino acid glutamic acid at position # 131 of SEQ ID No.:4. It is expected that the processing of human PMH-17 or PMH-18, in mature forms, potentially involves dimerization and elimination of the N-terminal region, in a manner analogous to the processing of the related protein TGF-β [Gentry et al, Molec & Cell. Biol., 8: 4162 (1988); Derynck et al. Nature, 316: 701 (1985)]. Therefore, it is contemplated that the mature, active species of human PMH-17 and PMH-18 may comprise homodimers of two polypeptide subunits where each subunit comprises amino acids from # 1 to # 224 of the SEQ ID No.:2 or from # 1 to # 231 of SEQ ID No .: 4, respectively, with a predicted molecular weight of approximately 49 to 52 kD. Additional active species containing at least amino acids from # 109 to # 211 of SEQ ID No.:2 or from # 116 to # 218 of SEQ ID No .:, thereby including the first and last residues of cysteine preserved. As with other members of the TGF-β / PMH protein family, the carboxyl terminal portion of the human PMH-17 and PMH-18 polypeptides exhibits greater sequence conservation than the amino terminal portion. Human PMH-17 (SEQ ID No.:l) or DNA sequence of PMH-18 (SEQ ID No: 3), or a portion thereof, can be used as a probe to identify a cell line human or tissue that synthesizes human PMH-17 or PMH-18, or a mRNA related to human PMH-17 or PMH-18. Briefly described, the RNA is extracted from a selected source of cells or tissue, and, or electrophoresed in an agarose gel with formaldehyde and transferred to nitrocellulose, or reacted with formaldehyde and applied as a stain. on nitrocellulose directly. The nitrocellulose is subsequently hybridized to a probe derived from the coding sequence of human PMH-17 and PMH-18. Alternatively, the sequence of human PMH-17 or PMH-18 can be used to design oligonucleotide primers that will specifically amplify a portion of human PMH-17 or PMH-18, or related coding sequences. It is contemplated that these primers derived from human PMH-17 and human PMH-18, would allow a person to specifically amplify the corresponding human PMH-17 or PMH-18, or related coding sequences, from mRNA templates, CDNA or genomic DNA. Once a positive source has been identified, by one of the methods described above, the mRNA is selected by chromatography on oligo (dT) cellulose and the cDNA is synthesized and cloned into the gtlO or other bacteriophage vectors? known by those skilled in the art, for example, the? ZAP, through established techniques (Toóle et al, supra). It is also possible to perform the amplification reaction directed by the oligonucleotide primer, described above, directly on a cDNA or pre-established human genomic library, which has been cloned into a bacteriophage vector?. In those cases, a library that produces a specifically amplified DNA product, which encodes a portion of the human PMH-17 or PMH-18 or related protein, could be subjected directly to selective screening, using the PMH-17 or PMH fragment -18, human, or related, amplified protein, which encodes DNA as a probe. Additional methods known to those skilled in the art can be used to isolate other full-length cDNAs, which encode human PMH-17 or PMH-18, or related proteins, or full-length cDNAs, encoding PMH -17 or PMH-18 or related proteins, of the invention, from species other than humans, particularly from other mammalian species.

EXAMPLE 2 BIOLOGICAL TESTS WITH CELLS W-20 A. Description of W-20 cells The use of bone marrow stromal cells, W-20, as an indicator cell line, is based on the conversion of these cells into cells similar to osteoblasts, after treatment with a PMH protein [Thies et al, Journal of Bone and Mineral Research, 5_: 305 (1990); and Thies et al, Endocrinology, 130: 1318 (1992)].

Specifically, W-20 cells are a clonal line of bone marrow stromal cells, derived from adult mice, by researchers from Dr. D.'s laboratory.

Nathan, Children's Hospital, Boston, MA. The treatment of W-20 cells with certain PMH proteins results in (1) an increased production of alkaline phosphatase, (2) the induction of cAMP-stimulated cAMP, and (3) the induction of osteocalcin synthesis by cells. Although (1) and (2) represent characteristics associated with the osteoblast phenotype, the ability to synthesize osteocalcin is a phenotypic property exhibited only by mature osteoblasts. In addition, to date it has been iSWiír? JSWSBffll "'observed the conversion of the W-20 stromal cells in cells similar to osteoblasts, only with the treatment with the PMH.In this way, the in vi tro activities, exhibited by the c & ks W-20 treated with PMH, correlate with the active bone formation, in vivo known for PMH.There are described two in vi tro tests useful in the comparison of the activities of the PMH, of novel osteoinductive molecules. B. Protocol of the Alkaline Fostase Assay in W-20 Cells.

W-20 cells are applied in 96 well tissue culture plates at a density of 10,000 cells per well in 200 μl media (DME with 10% calf fetal calf serum, 2 mM glutamine and 100 Units / milliliter of penicillin + 100 μg / ml streptomycin The cells were allowed to adhere overnight in an incubator at 37 ° C with 95% air and 5% C02. Each well, with a multi-channel pipettor, is replaced with an equal volume of the test sample placed in DME with 10% fetal calf serum, thermally inactivated, 2mM glutamine and 1% penicillin-streptomycin. The test samples were tested in triplicate.The test samples and standards were left in incubation for a period of 24 hours with the W-20 indicator cells.After 24 hours, the plates were removed from the incubator at 37 ° C. and the means of proof were removed from the cé Cells The W-2-U Ufe cell layers were washed 3 times with 200 μl per well of phosphate regulated salt solution, free of calcium and magnesium, and these washes were discarded. 50 μl of distilled water in glass was added to each well, and the test plates were then placed in a dry ice / ethanol bath to obtain a quick freeze. Once frozen, the test plates are removed from the dry ice / ethanol bath, and thawed at 37 ° C. This step is repeated 2 more times for a total of 3 freeze-thaw procedures. Once finished, the alkaline phosphatase bound to the membrane is available for measurement. 50 μl of the test mixture (50 mM glycine, 0.05% Triton X-100, 4 mM MgCl 2, 5 mM p-nitrophenol phosphate, pH = 10.3) were added to each test well and the plates The assays were then incubated for 30 minutes at 37 ° C in a shaking water bath at 60 oscillations per minute. At the end of the 30 minute incubation, the reaction was stopped by adding 100 μl of 0.2 N NaOH to each well and placing the test plates on ice. The spectrophotometric absorbance, for each well, was read at a wavelength of 405 nanometers. These values are then compared with known standards to give an estimate of the alkaline phosphatase activity in each sample. For example, using known amounts of p-nitrophenol phosphate, the absorbance values are generated. This is shown in Table I.

Table I Absorbanc values. The for standards > Known P-Nitrophenol Phosphate μmoles P-nitrophenol phosphate * '-Absorbance average (405 nm) 0.000 0 0.006 0.261 +/- .024 0.012 0.521 +/-. 031 0.018 0.797 +/-. 063 0.024 1.074 + /-.061 0.030 1.305 +/-. 083 The absorbance values for known amounts of PMH can be determined and converted to μmol of p-nitrophenol phosphate cleaved per unit time, as shown in Table II.

Table II Alkaline Phosphatase Values for Celalla W-20 Treatment with PMH-2 Reading Concentration of μmol 3S of substrate PMH-2 Absorbance Per hour ng / ml 405 nanometers 0 0.645 0. 024 1.56 0.696 0. 026 3.12 0.765 0. 029 6.25 0.923 0. 036 12.50 1.121 0. 044 25.0 1.457 0. 058 50.0 1.662 0. 067 100.0 1.977 0. 080 10 M ^ ^. »^ .. -" '' J &EL ':%' ^^ ^ »^? ~ M These values are then used to compare the activities of known amounts of PMH-17 and PMH-18 with PMH-2.

C. Radioimmunoassay Protocol (RIA) for Osteocalcin Plates, W-20 cells, were applied at a rate of 10 6 cells per well, in tissue culture dishes, from multiple wells, with 24 wells, in 2 ml of DME containing 10% fetal calf serum, thermally inactivated , 2mM glutamine. The cells were allowed to adhere overnight in an atmosphere of 95% air and 5% C02 at 37 ° C. The next day the medium was changed to DME containing 10% fetal calf serum, 2 mM glutamine and the test substance in a total volume of 2 ml. Each test substance was administered to the wells in triplicate. The test substances were incubated with W-20 cells, for a total of 96 hours, with replacement, at 48 hours, by the same test means. At the end of 96 hours, 50 μl of the test media from each well was removed and tested for osteocalcin production, using a radioimmunoassay for mouse osteocalcin. The details of the assay are described in the kit manufactured by Biomedical Technologies Inc., 378 Page Street, Stoughton, MA 02072. Reagents for the assay are found as product numbers BT-431 (mouse osteocalcin standard), BT-432 (Osteocalcin anti-mouse Cabra), BT-431R (osteocalcin iodine of mouse), BT-415 (normal goat serum) and BT-414 (IgG anti-goat donkey). The radioimmunoassay for os theocalcin synthesized by the W-20 cells, in response to treatment with PMH, is carried out as described in the protocol provided by the manufacturer. The values obtained for the test samples are compared with the values for known standards of mouse osteocalcin and with the amount of osteocalcin produced by the W-20 cells in response to the challenge with known amounts of PMH-2. The values for the synthesis of osteocalcin, induced by PMH-2, by the W-20 cells, is presented in Table III.

Table III Synthesis of Osteocalcin by Cells W-20 Concentration of PMH - 2 ng / ml Synthesis of osteocalcin ng / well 0 0.8 2 0.9 4 0.8 8 2.2 17 2.7 31 3.2 62 5.1 125 6.5 250 8.2 500 9.4 1000 10.0 15 fSitiSttil ^^^^ jfe - '^ aa & ^ ia ^^ EXAMPLE 3 SAMPATH-REDDI ESSAY MODIFIED BY ROSEN A modified version of the test for bone formation in rat, described in Sampath and Reddi, Proc. Nati Acad. Sci. United States of America, 80: 6591-6595 (1983), to evaluate the activity of PMH proteins, in bone and / or cartilage and / or other connective tissue. This modified test is called here, Sampath-Reddi test modified by Rosen. The step of ethanol precipitation, of the Sampath-Reddi process, is replaced by dialysis (if the composition is a solution) or by diafiltration (if the composition is a suspension) of the fraction to be tested, against water. The solution or suspension is then equilibrated at 0.1% TFA. The resulting solution is added to 20 mg rat matrix. A mimic sample of rat matrix, not treated with protein, serves as a control. The material is frozen and lyophilized and the resulting powder is enclosed in # 5 gelatin capsules. The capsules are implanted subcutaneously in the abdominal thoracic area of Long Evans rats, male, with an age of 21 to 49 days. The implants are removed after a period of 7 to 14 days. Half of each implant is used for analysis with alkaline phosphatase [See, Reddi et al, Proc. Nati Acad. Sci., 69: 1601 (1972)]. The other half of each implant is fixed and processed for histological analysis. Sections of 1 μm of glycol methacrylate are stained with Von Kossa and acid fuschine to give a score to the induced amount of bone and .it ^ f ^ as ^? u ^ cartilage, and other formation * of connective tissue .fi | pesent in each implant. The terms from +1 to +5 represent the area of each histological section of an implant, occupied by new bone and / or cartilage cells and the matrix. A score of +5 indicates that more than 50% of the implant is new bone and / or cartilage, produced as a direct result of the protein in the implant. A score of +4, +3, +2 and +1 indicates that more than 40%, 30%, 20% and 10% respectively of the implant, contains new cartilage and / or bone. Alternatively, the implants are inspected for the appearance of the tissue resembling the embryonic tendon, which is easily recognized by the presence of dense bundles of fibroblasts oriented in the same plane and tightly packed together. [Tendon / ligament-like tissue, is described, for example, in Ham and Cormack, Histoloqy (JB Lippincott Co. (1979), pp. 367-369, the disclosure of which is incorporated herein by reference.] These discoveries can be reproduced in additional tests in which tendon / ligament-like tissues are observed, in implants containing protein related to PMH-17 and PMH-18, proteins related to PMH-17 and PMH-18, This invention can be evaluated with respect to its activity in this assay.

EXAMPLE 4 Expression of PMH-17 and PMH-18 In order to produce murine, human, or other mammalian PMH-17 and PMH-18 proteins, the DNA encoding them, is transferred to an appropriate expression vector, and is introduced into the cells of the tilmiferous or other eukaryotic hosts or prokaryotic, preferred, through conventional techniques of genetic engineering. It is contemplated that the expression system "Preferred for human, recombinant, biologically active PMH-17 and PMH-18 is that of stably transformed mammalian cells. A person skilled in the art can construct mammalian expression vectors, employing the sequence of SEQ ID NO: SEQ ID No .: 3, or other DNA sequences encoding PMH-17 and PMH-18 or related proteins , or other modified sequences and known vectors, such as pCD [Okayama et al, Mol. Cell Biol., 2: 161 (1982), pJL3, pJL4 [Gough et al, EMBO J ^, 4: 645-653 (1985)] and pMT2 CXM. The mammalian expression vector pMT2 CXM is a derivative of p91023 (b) (Wong et al, Science 228: 810-815, 1985) which differs from the latter and which contains the ampicillin resistance gene, instead of the gene of resistance to tetracycline, and which also contains an XhoI site for the insertion of cDNA clones. The functional elements of pMT2 CXM have been described (Kaufman, RJ, 1985, Proc. Nati, Acad. Sci. United States of America 82 ^ 689-963) and include the VA adenovirus genes, the SV40 origin of replication including the enhancer 72 bp, the major late promoter of the adenovirus, which includes a 5 'splice site and most of the adenovirus tripartite guiding sequence, present in the late adenovirus mRNAs, a 3' splice acceptor site, an DHFR, the SV40 early polyadenylation site, (SV40) and the pBR322 sequences necessary for propagation in E. coli. The plasmid pMT2 CXM, is obtained by digestion with EcoRI of pMT2-VWF, which has been deposited in the American Type Culture Collection (ATCC), Rockville, MD (United States of America) under accession number ATCC 67122. Digestion with EcoRI cleaves the cDNA insert present in pMT2-VWF, producing pMT2 in linear form, which can be ligated and used to transform HB 101 or DH-5 from E. coli for ampicillin resistance. The plasmid DNA pMT2 can be prepared by conventional methods. The pMT2 CXM is then constructed using mutagenesis outside and inside the loop [Morinaga, et al, Biotechnology 8_4: 636 (1984)]. This eliminates the bases of 1075 to 1145 in relation to the Hind III site close to the SV40 origin of replication and the enhancer sequences of pMT2. In addition it inserts the following sequence: 5 'PO-CATGGGCAGCTCGAG-3' in nucleotide 1145. This sequence contains the recognition site for the restriction Xhol endonuclease. A derivative of pMT2 CXM, called pMT23, contains recognition sites for the restriction endonucleases PstI, Eco Rl, SalI and Xhol. Plasmid DNA pMT2 CXM and pMT23 can be prepared by conventional methods. The pEMC2β1 derived from pMT21 may also be convenient in the practice of the invention. PMT21 is derived from pMT2 which is derived from pMT2-VWF. As described above, Eco Rl digestion cleaves the cDNA insert present in pMT-VWF, producing pMT2 in the linear form, which can be ligated and used to transform HB S & amp; .4"101 or DH-5 of E. coli for resistance to the apicid.a.The DNA of plasmid pMT2 can be prepared by conventional methods.PMT21 is derived from pMT2 through the following two First, 76 bp is removed from the 5 'untranslated region of the DHFR cDNA that includes a 19-residue stretch of G from the G / C coleus for cloning the cDNA In this process, an Xhol site is inserted, to obtain the following sequence, immediately in the 5 'direction of the DHFR: 5' -CTGCAGGCGAGCCTGAATTCCTCGAGCCATCATG-3 'PstI Eco Rl Xhol Second, a single Clal site is introduced by digestion with EcoRV and Xbal, treatment with a Klenow fragment of DNA polymerase I and binding to a Clal linker (CATCGATG). This removes a 250 bp segment from the VAI region of the RNA associated with the adenovirus, but does not interfere with the expression or gene function of the VAI RNA. PMT21 is digested with Eco RlXhoI and used to derive vector PEMC2B1. A portion of the EMCV leader is obtained from pMT2-ECAT1 [S.K. Jung, et al, J. Virol 63: 1651-1660 (1989)] by digestion with EcoRI and PstI, resulting in a 2752 bp fragment. This fragment is digested with Taql producing a 508 bp EcoRI-Taql fragment which is purified by low melting point agarose gel electrophoresis. A 68 bp adapter and its complementary strand are synthesized with a 5 'Taql protruding end and a 3' Xhol protruding end having the following sequence: 5 'CGAGGTTAAAAAACGTCTAGGCCCCCCGAACCACGGGGACGTGGTTTTCCTTT Taql GAAAAACACGATTGC-3' Xhol This sequence matches the EMC virus guiding sequence, from nucleotide 763 to 827. It also changes the ATG at position 10 within the EMC virus leader to an ATT and is followed by an Xhol site. A three-way ligation of the EcoRI-16hoI fragment of pMT21, the EcoRI-Taql fragment of the EMC virus, and the Taqi-16hoI adapter of the 68 bp oligonucleotide adapter, results in the vector pEMC2ßl. This vector contains the SV40 origin of replication and enhancer, the main late promoter of the adenovirus, a copy of the cDNA of most of the tripartite leader sequence of the adenovirus, a small hybrid intervening sequence, a SV40 polyadenylation signal and the VAI gene of the adenoviruses, DHFR and β-lactamase markers and an EMC sequence, in appropriate ratios to direct the high level expression of the desired cDNA in mammalian cells. The construction of vectors may involve the modification of DNA sequences related to PMH-17 and PMH-18. For example, the cDNA of PMH-17 and PMH-18 can be modified by removing the non-coding nucleotides at the ends of the coding region. The non-coding nucleotides, deleted, may or may not be replaced by other sequences known to be beneficial for expression. These vectors are transformed into host cells suitable for the expression of the proteins related to PMH-17 and PMH-18. Additionally, the sequence of SEQ ID No .: 1, SEQ ID 5 No .: 3 or other sequences that encode the proteins related to PMH-17 and PMH-18 can be manipulated to express a protein PMH-17 or PMH-18, mature, or related protein, eliminating the sequences of propeptides that encode PMH-17 or PMH-18 and replacing with sequences that encode the complete propeptides of other PMH proteins. A person skilled in the art can manipulate the sequences of SEQ ID No.:l or SEQ ID No .: 3, eliminating or replacing the regulatory sequences of mammals, flanking the coding sequence, with bacterial sequences, to create bacterial vectors for intracellular or extracellular expression by bacterial cells. For example, the coding sequences could be further manipulated (for example, bound to other known or modified linkers by removing non-coding sequences thereof or by altering the nucleotides found therein, by other known techniques). The coding sequence related to PMH-17 or PMH-18, modified, could Then inserted into a known bacterial vector, using methods such as those described in T. Taniguchi et al, Proc. Nati Acad. Sci. United States of America, 77: 5230-5233 (1980). This exemplary bacterial vector could then be transformed into host cells bacterial and the protein related to PMH-17 and PMH-18 - ^^ s' could be expressed by the same. One to produce the xtracellular expression of proteins related to PMH-17 or PMH-18, in bacterial cells, can be seen, for example, in the European patent application EPA 177,343. Similar manipulations for the construction of an insect vector can be carried out [see, for example, the procedures described in published European patent application, 155,476] for expression in insect cells. A yeast vector could also be constructed using yeast regulatory sequences, for the intracellular or extracellular expression of the factors of the present invention, by the yeast cells. [See, for example, the procedures described in published PCT application WO86 / 00639 and European patent application 123,289]. A method for producing high levels of a PMH-17 or PMH-18 protein, or related proteins, of the invention, in mammalian cells, may involve the construction of cells containing multiple copies of the heterologous PMH-17 or PMH gene. -18. The heterologous gene is linked to an amplifiable marker, for example the hydrofolate reductase gene (DHFR) for which, cells containing the increased gene copies, can be selected for propagation at increasing concentrations of methotrexate (MTX) according to with the procedures of Kaufman and Sharp, J. Mol. Biol., 159: 601-629 (1982). This approach can be employed with a number of different cell types. For example, a plasmid containing a DNA sequence for a protein PMH-17 or PMH-18 or related proteins of the invention, in functional association with other sequences of the plasmid that allow the expression thereof and the expression plasmid pAdA26SV (A ) 3 of the DHFR [Kaufman and Sharp, Mol. Cell. Biol., 2: 1304 (1982)] can be introduced together in CHO cells deficient in DHFR, DUKX-BII, through several methods including co-precipitation with calcium phosphate and transfection, electroporation or fusion to the protoplast. Transformants expressing DHFR are selected for culture in alpha media with dialyzed calf fetal serum, and are subsequently selected for amplification by growing in increasing concentrations of MTX (eg, sequential steps in 0.02, 0.2, 1.0 and 5). μM of MTX) as described in Kaufman et al, J. Mol. Biol., 5: 1650 (1983). The transformants are cloned, and the expression of the PMH-17 or biologically active PMH-18 is inspected by the rat bone formation test, Sampath-Reddi modified by Rosen, described above in example 3. The expression of the protein PMH-17 and PMH-18 should be increased with increasing levels of resistance to MTX. The PMH-17 and PMH-18 polypeptides are characterized using standard procedures known in the art, such as pulse labeling with [35S] raetiomna or cysteine and polyacrylamide gel electrophoresis. Similar procedures can be followed to produce other related PMH-17 and PMH-18 proteins, or related proteins.

EXAMPLE 5 Biological Activity of PMH-17 and PMH-18 expressed To measure the biological activity of the expressed PMH-17 and PMH-18, or related proteins obtained in example 4 above, the proteins are recovered from cell culture and purified by isolating PMH-17, PMH-18 or related proteins, of other proteinaceous materials with which they are produced together, as well as other contaminants. The purified protein can be tested according to the rat bone formation assay described in Example 3. The purification is carried out using standard procedures known to those skilled in the art. The protein analysis is carried out using standard techniques such as SDS-PAGE acrylamide [Laemmli, Nature 227: 680 (1970)] stained with silver [Oakley, et al. Anal.

Biochem. 105: 361 (1980)] and by immunoabsorption [To bin, et al. Proc. Nati Acad. United States of America 76: 350 (1979)].

EXAMPLE 6 Using the Northern analysis can analyze the proteins PMH-17 and PMH-18, and related proteins, with respect to their effects in several cell lines. Suitable cell lines include the cell lines derived from the protrusions of the extremities of the E13 mouse.

After 10 days of trimerizing with PMH-17, PMH-18 or related proteins, the cell PhiPype is examined histologically for evidence of tissue differentiation. In addition, North'ern analysis of the mRNA of cells treated with protein PMH-17, PMH-18 or related proteins, can be carried out for several markers, including one or more of the following markers for bone, cartilage and / or tendon / ligament, as described in Table IV: Table IV Marker Bone Cartilage Tendon / Ligament Osteocalcin + Phosphatase + Alkaline Protein +/- 1 Nucleus of Proteoglycan Collagen Type + I Collagen Type +/- 1 II Decorin + + + Elastin +/- 3 9 + 1-The marker is seen early, the marker is not seen as mature forms of bone tissue. 2-The marker depends on the site of the tendon; It is the strongest in the bone interface. 3-The marker is seen at low levels.

EXAMPLE 7 Essay of Embryonic Pluripotent Cells In order to analyze the effects of the PMH-17 and PMH-18 proteins of the present invention, it is possible to test the effects of growth and differentiation in vi tro on a number of available embryonic pluripotent cell lines. One of those cell lines is the ES-E14TG2, which is available from the American Type Culture Collection in Rockville, MD. In order to carry out the assay, the cells can be propagated in the presence of 100 units of LIF to maintain them in an undifferentiated state. The tests are established by first removing the LIF and adding the cells in suspension, in what is known as embroid bodies. After 3 days the embryo bodies were applied on gelatin coated plates (12-well plates for PCR analysis and 24-well plates for immunocytochemistry) and treated with the proteins to be tested. The cells are supplied with nutrients and treated with the protein factor every 2 - 3 days. The cells can be adapted in such a way that the assays can be carried out in media supplemented with 15% Bovine Fetal Serum (FBS) or with media defined by CDM containing much lower amounts of FBS. At the end of the treatment period (which varied from 7 to 21 days) the RNA is collected from the cells and analyzed by the multiplex, quantitative Polymerase Chain Reaction (PCR) for the following markers: Brachyury, a mesodermal marker , AP-2, an ectodermal marker, and HNF-3 to an endodermal marker. Through the immunocytochemistry, táttfj-én it is possible to detect the differentiation of neuronal cells (glia and neurons), muscle cells (cardiomyocytes, skeletal and smooth muscle) and several other phenotypic ions such as proteoglycan core protein (cartilage), and cytokeratins (epidermis). Since these cells have a tendency to differentiate autonomously when the LIF is removed, the results are always quantified by comparison with an untreated control. The foregoing descriptions detail currently preferred embodiments of the present invention. It is expected that those skilled in the art will come up with numerous modifications and variations in the practice thereof, when considering these descriptions. It is believed that those modifications and variations are contained within the claims appended hereto.

LIST OF SEQUENCES (1) GENERAL INFORMATION: i) APPLICANT: Genetics Institute, Inc. ri- (ii) TITLE OF THE INVENTION: Bone Morphogenetic Protein (BMP) -17 and BMP-18 Combinations. (iii) NUMBER OF SEQUENCES: 4 (iv) ADDRESS FOR CORRESPONDENCE: (A) SENDER: GENETICS INSTITUTE, INC. (B) STREET: 87 CAMBRIDGEPARK DRIVE (C) CITY: CAMBRIDGE (D) STATE: Massachusetts (E) COUNTRY: USA (F) POSTAL CODE: 02140 (v) COMPUTER LEADABLE FORM: (A) TYPE OF MEDIA: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS / MS-DOS (D) SOFTWARE: Patentln Relay # 1.0, Version # 1.30 (vi) CURRENT REQUEST DATA: (A) NUMBER OF APPLICATION: PCT / (B) DATE OF PRESENTATION: with this (C) CLASSIFICATION: (viii) INFORMATION OF THE POWDER / AGENT: (A) NAME: Lazar, Steven R. (B) REGISTRATION NUMBER: 32,618 (C) NUMBER OF RER / FILE: Gl 5307-PCT (x) INFORMATION FOR TELECOMMUNICATIONS: (A) TELEPHONE: (617) 498-8260 (B) TELEFAX: (617) 876-8581 (2) INFORMATION FOR SEQ ID NO.:l: ( i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 1101 base pairs (B) TYPE: nucleic acid (C) CHAIN TYPE: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: cDNA (ix) CHARACTERISTICS: (A) NAME / KEY: pro_peptide (B) LOCATION: 1. 426 6?TERISTICS: ^ (A) NAME / CLA ^ í mat_peptide (B) LOCATION: 427..1098 (ix) CHARACTERISTICS: (A) NAME / KEY: CDS (B) LOCATION: 1..1098 (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: l; ATG CAG CCC CTG TGG CTC TGC TGG GCA CTC TGG GTG TTG CCC CTG GCC 48 Met Gln Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala -142 -140 -135 -130 AGC CCC GGG GCC GCC CTG ACC GGG GAG CAG CTC CTG GGC AGC CTG CTG 96 Ser Pro Gly Ala Ala Leu Thr Gly Glu Gln Leu Leu Gly Ser Leu Leu -125 -120 -115 CGG CAG CTG CAG CTC AAA GAG GTG CCC ACC CTG GAC AGG GCC GAC ATG 144 Arg Gln Leu Gln Leu Lys Glu Val Pro Thr Leu Asp Arg Wing Asp Met -110 -105 -100 -95 GAG GAG CTG GTC ATC CCC ACC CAC GTG AGG GCC CAG TAC GTG GCC CTG 192 Glu Glu Leu Val He Pro Thr His Val Arg Ala Gln Tyr Val Ala Leu -90 -85 -80 CTG CAG CGC AGC CAC GGG GAC CGC TCC CGC GGA AAG AGG TTC AGC CAG 240 Leu Gln Arg Ser His Gly Asp Arg Ser Arg Gly Lys Arg Phe Ser Gln -75 -70 -65 AGC TTC CGA GAG GTG GCC GGC AGG TTC CTG GCG TTG GAG GCC AGC ACA 288 Ser Phe Arg Glu Val Wing Gly Arg Phe Leu Wing Leu Glu Wing Ser Thr -60 -55 -50 CAC CTG CTG GTG TTC GGC ATG GAG CAG CGG CTG CCG CCC AAC AGC GAG 336 His Leu Leu Val Phe Gly Met Glu Gln Arg Le u Pro Pro Asn Ser Glu -45 -40 -35 CTG GTG CAG GCC GTG CTG CGG CTC TTC CAG GAG CCG GTC CCC AAG GCC 384 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala -30 -25 - 20 -15 GCG CTG CAC AGG CAC GGG CGG CTG TCC CCG CGC AGC GCC CGG GCC CGG 432 Ala Leu His Arg His Gly Arg Leu Ser Pro Arg Ser Ala Arg Ala Arg -10 -5 1 GTG ACC GTC GAG TGG CTG CGC GTC CGC GAC GAC GGC CC AAC CGC ACC 480 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 5 10 15 TCC CTC ATC GAC TCC AGG CTG GTG TCC GTC CAC GAG AGC GGC TGG AAG 528 Ser Leu He Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 20 25 30 GCC TTC GAC GTG ACC GAG GCC GTG A &? - TTC TGG CAG CAG CTG AGC CGG 576 Wing Phe Asp Val Thr Glu Wing vl Asn Phe Trp Gln Gln Leu Ser Arg 35 40 45 50 CCC CGG CAG CCG CTG CTG CTA CAG GTG TCG GTG CAG AGG GAG CAT CTG 624 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 55 60 65 GGC CCG CTG GCG TCC GGC GCC CAC AAG CTG GTC CGC TTT GCC TCG CAG 672 Gly Pro Leu Wing Ser Gly Wing His Lys Leu Val Arg Phe Wing Ser Gln 70 75 80 GGG GCG CCA GCC GGG CTT GGG GAG CCC CAG CTG GAG CTG CAC ACC CTG 720 Gly Wing Pro Wing Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 85 90 95 GAC CTT GGG GAT TAT GGA GCT CAG GGC GAC TGT GAC CCT GAA GCA CCA 768 Asp Leu Gly Asp Tyr Gly Wing Gln Gly Asp Cys Asp Pro Glu Wing Pro 100 105 110 ATG ACC GAG GGC ACC CGC TGC TGC CGC CAG GAG ATG TAC ATT GAC CTG 816 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr He Asp Leu 115 120 125 130 CAG GGG ATG AAG TGG GCC GAG AAC GTG CTG GAG CCC CCG GGC TTC 864 Gln Gly Met Lys Trp Wing Glu Asn Trp Val Leu Glu Pro Pro Gly Phe 135 140 145 CTG GCT TAT GAG TGT GTG GGC ACC TGC CGG CAG CCC CCG G AG GCC CTG 912 Leu Ala Tyr Glu Cys Val Gly Thr Cys Arg Gln Pro Pro Glu Ala Leu 150 155 160 GCC TTC AAG TGG CCG TTT CTG GGG CCT CGA CAG TGC ATC GCC TCG GAG 960 Wing Phe Lys Trp Pro Phe Leu Gly Pro Arg Gln Cys He Wing Ser Glu 165 170 175 ACT GCC TCG CTG CCC ATG ATC GTC AGC ATC AAG GAG GGA GGC AGG ACC 1008 Thr Wing Ser Leu Pro Met He Val Ser He Lys Glu Gly Gly Arg Thr 180 185 190 AGG CCC CAG GTG GTC AGC CTG CCC AAC ATG AGG GTG CAG AAG TGC AGC 1056 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 195 200 205 210 TGT GCC TCG GAT GGT GCG CTC GTG CCA AGG AGG CTC CAG CCA 1098 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro 215 220 TAG_1101_(2) INFORMATION FOR SEQ ID N0.:2: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 366 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: Protein (xi) ) SEQUENCE DESCRIPTION: SEQ ID NO: 2: Met Gln Pro Leu Trp Leu Cys Trp Wing Leu Trp Val Leu Pro Leu Wing -142 -140 -135 -130 Ser Pro Gly Ala Wing Leu Thr Gly Gllu Gln Leu Leu Gly Ser Leu Leu -125 -120 -115 Arg Gln Leu Gln Leu Lys Glu Val Pro Thr Leu Asp Arg Ala Asp Met -110 -105 -100 -95 Glu Glu Leu Val He Pro Thr His Val Arg Ala Gln Tyr Val Ala Leu -90 -85 -80 Leu Gln Arg Ser His Gly Asp Arg Ser Arg Gly Lys Arg Phe Ser Gln -75 -70 -65 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Wing Leu Glu Wing Being Thr -60 -55 -50 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu -45 -40 -35 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala -30 -25 -20 -15 Ala Leu His Arg His Gly Arg Leu Ser Pro Arg Be Ala Arg Ala Arg -10 -5 1 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 5 10 15 Ser Leu He Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 20 25 30 Wing Phe Asp Val Thr Glu Wing Val Asn Phe Trp Gln Gln Leu Ser Arg 35 40 45 50 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 55 60 65 Gly Pro Leu Wing Ser Gly Wing His Lys Leu Val Arg Phe Wing Ser Gln 70 75 80 Gly Wing Pro Wing Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 85 90 95 Asp Leu Gly Asp Tyr Gly Wing Gln Gly Asp Cys Asp Pro Glu Ala Pro 100 105 110 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr He Asp Leu 115 120 125 130 Gln Gly Met Lys Trp Wing Glu Asn Trp Val Leu Glu Pro Pro Gly Phe 135 140 145 Leu Ala Tyr Glu Cys Val Gly Thr Cys Arg Gln Pro Pro Glu Ala Leu 150 155 160 Wing Phe Lys Trp Pro Phe Leu Gly Pro Arg Gln Cys He Wing Ser Glu 165 170 175 Thr Wing Ser Leu Pro Met He Val Ser He Lys Glu Gly Gly Arg Thr 180 185 190 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 195 200 205 210 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro 215 220 (2) INFORMATION FOR SEQ ID NO.:3: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 1101 base pairs (B) TYPE: nucleic acid (C) TYPE OF CHAIN: simple (D) TOPOLOGY: linear (n) TYPE OF MOLECULE: DNA (genomic) (ix) CHARACTERISTICS: (A) NAME / KEY: pro_peptide (B) LOCATION: 1.405 (ix) CHARACTERISTICS: (A) NAME / KEY: matjpeptide (B) LOCATION: 406..1098 (ix) FEATURE: (A) NAME / KEY: CDS (B) LOCATION: 1..1098 (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 3: ATG TGG CCC CTG TGG CTC TGC TGG GCA CTC TGG GTG CTG CCC CTG GCT 48 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala -135 -130 -125 -120 GGC CCC GGG GCG GCC CTG ACC GAG GAG CAG CTC CTG GGC AGC CTG CTG 96 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu -115 -110 -105 ÜÉ CGG CAG CTG CAG CTC AGC GAG GTG CCC GTA CTG GAC AGG GCC GAC ATG 144 Arg Gln Leu Gln Leu Ser Glu ^ al Pro Val Leu Asp Arg Ala Asp Met -100 -95 -90 GAG AAG CTG GTC ATC CCC GCC CAC GTG AGG GCC CAG TAT GTA GTC CTG 192 Glu Lys Leu Val He Pro Wing His Val Arg Wing Gln Tyr Val Val Leu -85 -80 -75 CTG CGG CGC AGC CAC GGG GAC CGC rcc > CGC GGA AAG AGG TTC AGC CAG 240 Leu Arg Arg Ser His Gly Asp Arg Se'r Arg Gly Lys Arg Phe Ser Gln -70 -65 -60 AGC TTC CGA GAG GTG GCC GGC AGG TC CTG GCG TCG GAG GCC AGC ACÁ 288 Ser Phe Arg Glu Val Wing Gly Arg Phe Leu Wing Ser Glu Wing Ser Thr -55 -50 -45 -40 CAC CTG CTG GTG TTC GGC ATG GAG CAG CGG CTG CCG CCC AAC AGC GAG 336 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu -35 -30 -25 CTG GTG CAG GCC GTG CTG CGG CTC TTC CAG GAG CCG GTC CCC AAG GCC 384 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Wing -20 -15 -10 GCG CTG CAC AGG CAC GGG CGG CTG TCC CCG CGC AGC GCC CAG GCC CGG 432 Ala Leu His Arg His Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg -5 1 5 GTG ACC GTC GG TGG CTG CGC GTC CGC GAC GAC GGC TCC AAC CGC ACC 480 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 10 15 20 25 TCC CTC ATC GAC TCC AGG CTG GTG TCC GTC CAC GAG AGC GGC TGG AAG 528 Ser Leu He Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 30 35 40 GCC TTC GAC GTG ACC GAG GCC GTG AAC TTC TGG CAG CAG CTG AGC CGG 576 Wing Phe Asp Val Thr Glu Wing Val Asn Phe Trp Gln Gln Leu Ser Arg 45 50 55 CCC CGG CAG CCG CTG CTG CTA CAG GTG TCG GTG CAG AGG GAG CAT CTG 624 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 60 65 70 GGC CCG CTG GCG TCC GGC GCC CAC AAG CTG GTC CGC TTT GCC TCG CAG 672 Gly Pro Leu Wing Ser Gly Wing His Lys Leu Val Arg Phe Wing Ser Gln 75 80 85 GGG GCG CCA GCC GGG CTT GGG GAG CCC CAG CTG GAG CTG CAC ACC CTG 720o Wing Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 90 95 100 105 GAC CTC AGG GAC TAT GGA GCT CAG GGC GAC TGT GAC CCT GAA GCA CCA 768 Asp Leu Arg Asp Tyr Gly Wing Gln Gly Asp Cys Asp Pro Glu Ala Pro 110 115 120 ATG ACC GAG GGC ACC CGC TGC TGC CGC CAG GAG ATG TAC ATT GAC CTG 816 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr He Asp Leu 125 130. 135 CAG GGG ATG AAG TGG GCC AAG AAC TGG GTG CTG GAG CCC CCG GGC TTC 864 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 140 145 150 CTG GCT TAC GAG TGT GTG GGC ACC TGC CAG CAG CCC CCG GAG GCC CTG 912 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 155 160 165 GCC TTC AAT TGG CCA TTT CTG GGG CCG CGA CAG TGT ATC GCC TCG GAG 960 Wing Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys He Wing Ser Glu 170 175 180 185 ACT GCC TCG CTG CCC ATG ATC GTC AGC ATC AAG GAG GGA GGC AGG ACC 1008 Thr Wing Ser Leu Pro Met He Val Ser He Lys Glu Gly Gly Arg Thr 190 195 200 AGG CCC CAG GTG GTC AGC CTG CCC AAC ATG AGG GTG CAG AAG TGC AGC 1056 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 205 210 215 TGT GCC TCG GAT GGG GCG CTC GTG CCA AGG AGG CTC CAG CCA 1098 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro 220 225 230 TAG_1101_(2) INFORMATION FOR SEQ ID NO.:4: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 366 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: Protein (xi) ) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: Met Trp Pro Leu Trp Leu Cys Trp Wing Leu Trp Val Leu Pro Leu Wing -135 -130 -125 -120 Gly Pro Gly Wing Wing Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu -115 -110 -105 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Wing Asp Met -100 -95 -90 Glu Lys Leu Val He Pro Wing His Val Arg Wing Gln Tyr Val Val Leu -85 -80 -75 Leu Arg Arg Ser His Gly Asp Arg Be Arg Gly Lys Arg Phe Ser Gln -70 -65 -60 Be Phe Arg Glu Val Wing Gly Arg Phe Leu Wing Ser Glu Wing Ser Thr -55 -50 -45 -40 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu -35 -30 -25 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Wing -20 -15 -10 Wing Leu His Arg His Gly Arg Leu Ser Pro Arg Wing Wing Gln Wing Arg -5 1 5 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 10 15 20 25 Ser Leu He Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 30 35 40 Wing Phe Asp Val Thr Glu Wing Val Asn Phe Trp Gln Gln Leu Ser Arg 45 50 55 Pro Arg Gln Pro Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 60 65 70 Gly Pro Leu Wing Ser Gly Wing His Lys Leu Val Arg Phe Ala Ser Gln 75 80 85 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 90 95 100 105 Asp Leu Arg Asp Tyr Gly Wing Gln Gly Asp Cys Asp Pro Glu Wing Pro 110 115 120 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr He Asp Leu 125 130 135 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 140 145 150 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 155 160 165 Wing Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys He Wing Ser Glu 170 175 180 185 Thr Ala Ser Leu Pro Met He Val Ser He Lys Glu Gly Gly Arg Thr 190 195 200 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 205 210 215 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro 220 225 230

Claims

1. An isolated DNA molecule, comprising a DNA sequence, characterized in that it is selected from the group consisting of: (a) nucleotides # 1, 232, 406, 427, 751, or 796 to # 1059 or 1098 of SEQ ID No.:l; (b) nucleotides encoding amino acids # -142, -65, -7, 1, 109 or 124 to # 211 or 224 of SEQ ID No .: 2; (c) nucleotides # 1, 232, 406, 427, 751, or 796 to # 1059 or 1098 of SEQ ID No.:3; (d) the nucleotides encoding amino acids # -135, -58, 1, 8, 116 or 131 to # 218 or 231 of SEQ ID No.:4; and (e) human, natural allelic sequences, and codon sequences, degenerate, equivalent, of (a), (b), (c) or (d).

2. A host cell, characterized in that it is transformed with the DNA sequence according to claim 1.

3. A vector characterized in that it comprises a DNA molecule according to claim 1, in functional association with an expression control sequence for the same.

4. A host cell characterized in that it is transformed with a vector according to claim 3.

5. An isolated DNA molecule, characterized in that it comprises a DNA sequence consisting of the nucleotides of # 427 to # 1098 of SEQ ID No.:l.

6. An isolated DNA molecule, characterized in that it comprises a DNA sequence consisting of the nucleotides of # 406 to # 1098 of SEQ ID NO: 3.

7. A vector characterized in that it comprises a DNA molecule according to claim 5, in functional association with an expression control sequence for the same.

8. A host cell characterized in that it is transformed with the vector according to claim 7.

9. A vector characterized in that it comprises a DNA molecule according to claim 6, in functional association with an expression control sequence for the same.

10. A host cell characterized in that it is transformed with the control vector according to claim 9.

11. A method to produce a bone morphogenetic protein, human, purified, PMH-17 and 'OR ?.. PMH-18, the method is characterized in that it comprises the steps of: (a) culturing a host cell transformed with a DNA molecule according to claim 1; Y (b) recovering and purifying the human PMH-17 and PMH-18 proteins from the culture medium.

12. The method according to claim 11, characterized in that the host cell is transformed with a DNA molecule comprising a DNA coding sequence consisting of nucleotide # 427 to # 1098 of SEQ ID No.:l.

13. The method according to claim 12, characterized in that the host cell is a mammalian cell and the DNA molecule further comprises a DNA sequence encoding a propeptide from a member of the TGF-β superfamily of proteins, the DNA sequence encodes a propeptide that is linked, in an appropriate reading frame, to the coding sequence of the DNA.

14. The method according to claim 11, characterized in that the host cell is transformed with a DNA molecule comprising a DNA coding sequence consisting of the nucleotides of # 406 to # 1098 of SEQ ID NO: 3.

15, The method according to claim 14, characterized in that the host cell is a mammalian cell and the DNA molecule further comprises a DNA sequence encoding a propeptide from a member of the TGF-β superfamily of proteins, the sequence of DNA encoding a propeptide is linked, in an appropriate reading frame, to the coding sequence of the DNA.

16. A polypeptide of the bone morphogenetic protein 17, purified, characterized in that it comprises the amino acid sequence of the amino acid from # 1 to # 224 which is presented in SEQ ID No.:2.

17. A purified PMH-17 polypeptide, according to claim 16, characterized in that the polypeptide is a dimer and that at least one subunit comprises the amino acid sequence of amino acid # 1 to # 224 of SEQ ID NO: 2.

18. A polypeptide of the bone morphogenetic protein 17, purified, produced by the steps of: (a) culturing a transformed cell with a DNA molecule according to claim 5; and (b) recovering and purifying, from the culture medium, a polypeptide comprising the amino acid sequence of amino acid # 1 to # 224 of SEQ ID No.:2.

19. A polypeptide of bone morphogenetic protein 17, purified, according to claim 18, characterized in that the polypeptide is a dimer comprising two subunits, wherein one subunit comprises the amino acid sequence of amino acid # 1 to # 224 of the SEQ ID No.:2, and a subunit comprises an amino acid sequence for a bone morphogenetic protein, selected from the group consisting of PMH-1, PMH-2, PMH-3, PMH-4, PMH-5, PHM-6, PMH-7, PMH-8, PMH-9, PMH-10, PMH-11, PMH-12, PMH-13, PMH-15 and PMH-16.

20. A polypeptide of bone morphogenetic protein 18, purified, characterized in that it comprises the amino acid sequence of amino acid # 1 to # 231 as presented in SEQ ID No.:4.

21. A purified PMH-18 polypeptide according to claim 20, characterized in that the polypeptide is a dimer and that at least one subunit comprises the amino acid sequence of amino acid # 1 to # 231 of SEQ ID NO: 4.

22. A polypeptide of the bone morphogenetic protein 18, purified, characterized in that it is produced by the steps of: (a) culturing a transformed cell with a DNA molecule, according to claim 6; and (b) recovering and purifying, from the culture medium, a polypeptide comprising the amino acid sequence of amino acid # 1 to amino acid # 231 of SEQ ID No.:4.

23. A polypeptide of bone morphogenetic protein 18, purified, according to claim 22, characterized in that the polypeptide is a dimer comprising 2 subunits, wherein one subunit comprises the amino acid sequence of amino acid # 1 to amino acid # 231 of SEQ ID No .: 4, and a subunit comprises an amino acid sequence for a bone morphogenetic protein, selected from the group consisting of PMH-1, PMH-2, PMH-3, PMH-4, PMH-5, PHM-6, PMH-7, PMH-8, PMH-9, PMH-10, PMH-11, PMH-12, PMH-13, PMH-15 and PMH-16.

24. A chimeric DNA molecule, characterized in that it comprises a DNA sequence encoding a propeptide from a member of the TGF-β protein superfamily bound, in frame, to a DNA sequence encoding a bone morphogenetic protein polypeptide. , the PMH-17 polypeptide comprises amino acids from # 1 to # 224 of SEQ ID NO: 2.

25. A chimeric DNA molecule characterized in that it comprises a DNA sequence encoding a propeptide from a member of the TGF-β protein superfamily, linked, in frame, to a DNA sequence encoding a bone morphogenetic protein polypeptide. , the PMH-18 polypeptide comprises of amino acid # 1 to # 231 of SEQ ID NO: 4.

26. Antibodies, characterized in that they are for a purified PMH-17 protein, in accordance with claim 16. HgÜ X

27. Antibodies, characterized in that they are for a purified PMH-18 protein, according to claim 20. 10 fifteen twenty 25 30