CN1661070A - Relationship between angiotensin I converting enzyme gene and essential hypertension - Google Patents

Abstract

The invention discloses a method for detecting the susceptibility of essential hypertension, which includes detecting whether there is a variation in the angiotensin I converting enzyme gene ACE, transcript and/or protein of the individual compared with the normal, and the existence of the variation indicates that the Individuals are more likely to suffer from essential hypertension than the normal population. The invention also discloses a corresponding detection kit.

Description

Relationship between angiotensin I converting enzyme gene and essential hypertension

technical field

The present invention relates to the fields of molecular biology and medicine. More specifically, it involves the single nucleotide polymorphism (singlenucleotide polymorphism, SNP) of angiotensin I converting enzyme gene (angiotensin I converting enzyme, ACE) and its correlation with essential hypertension. The present invention also relates to methods and kits for detecting these SNPs.

Background technique

Hypertension refers to a group of clinical syndromes with elevated systolic or diastolic blood pressure. There is an obvious causal relationship between the increase of blood pressure and the occurrence of coronary heart disease, renal dysfunction, hypertensive heart disease and hypertension complicated with stroke. The latest diagnostic criteria for hypertension are systolic blood pressure ≥ 19kpa (140mmHg) or diastolic blood pressure ≥ 12kpa (90mmHg), and those who meet one of them can be diagnosed as hypertension.

Most hypertensive patients only have mild subjective symptoms in the early stage of high blood pressure, such as headache, dizziness, insomnia, tinnitus, irritability, difficulty in concentrating work and study, and fatigue. With the development of the disease, especially when complications occur, the symptoms gradually increase and become more obvious, such as numbness and stiffness of fingers, pain in the lower limbs when walking more, or muscle pain and tension in the neck and back. When there is palpitation, shortness of breath, chest tightness, and precordial pain, it indicates that the heart has been involved. When there is nocturnal frequent urination, polyuria, and light urine, it indicates that the kidneys are involved and the renal arterioles are hardened. If a hypertensive patient suddenly develops confusion, deep and irregular breathing, and incontinence, etc., it may indicate cerebral hemorrhage. If one side of the body gradually becomes inconvenient, numb or even paralyzed, it should be suspected whether there is a cerebral thrombosis.

There were no obvious abnormal signs in the early stage of hypertension. Abnormal signs may appear when the brain, heart, kidney and other vital organs are slightly damaged. Common heart abnormalities include leftward shift of the apical beat, lifting-like pulsation sensation in the precordial area, enhanced first heart sound in the apical area, enhanced second heart sound in the aortic valve area, and systolic murmurs and diastolic murmurs. Arteriosclerosis and left ventricular hypertrophy, and a galloping rhythm in the apex may indicate heart failure. In addition, earlobe creases, capillary pulsation, hard or pulseless radial artery, and intermittent claudication of the lower extremities are also common.

In addition, due to certain predisposing factors or the development of hypertension itself, some hypertensive patients may have a significant or sudden increase in blood pressure, accompanied by damage to the brain, heart, kidney, retina and other important organs, which is seriously life-threatening, and a series of Special clinical signs are called hypertensive emergencies. The incidence of hypertensive emergencies accounts for 5% of the hypertensive population, common hypertensive encephalopathy, cerebral hemorrhage, acute left heart failure, clonidine acute withdrawal syndrome, acute myocardial infarction, rapidly progressive malignant hypertension, etc.

About 5% of hypertensive patients have no symptoms, do not know when blood pressure rises, and do not know when complications of blood vessel and organ damage have occurred. have high blood pressure. Therefore, finding out the genetic cause of hypertension, early prevention and detection can effectively control the incidence of hypertension and minimize the damage of the disease to the human body.

Essential hypertension (essential hypertension, EH), also called hypertension, is an independent disease with its own etiology, occurrence, development, outcome, and clinical manifestations. Clinically, it is mainly manifested as an increase in arterial blood pressure. Accounting for more than 90% of hypertensive patients in the population, the pathogenesis is not fully understood at present, and it is mainly diagnosed as essential hypertension (hypertension) only after the hypertension caused by other diseases has been ruled out. The increase in arterial blood pressure is mainly due to the increase in the resistance of peripheral small arteries, and at the same time there are varying degrees of increase in blood volume and cardiac output. In the late stage, the heart, brain, kidney and other organs are often involved, and serious complications such as hypertension, heart disease, heart failure, renal dysfunction, and cerebral hemorrhage occur. The treatment of essential hypertension is mainly to lower blood pressure while preventing the occurrence of complications. The causes of death in patients with essential hypertension are cerebrovascular accident, cardiovascular accident and renal insufficiency. In my country, cerebrovascular accident is more common, followed by heart failure and uremia. In European and American countries, heart failure is more common, and cerebrovascular accident is more common. and uremia followed.

As the most common cardiovascular disease, the incidence of essential hypertension is increasing year by year, and it can cause serious heart, brain, and kidney complications. It is a major risk factor for stroke and coronary heart disease. EH is a polygenic disease caused by the interaction of genetic factors and environmental factors. Finding EH-related genes and clarifying the genetic mechanism of hypertension has become a research hotspot.

Although there have been many studies on various gene polymorphisms and essential hypertension, there is no report confirming the correlation between ACE gene and essential hypertension, and there is no confirmation that the SNP of ACE gene is associated with essential hypertension sex reports.

To sum up, in order to finally realize the treatment of hypertension, there is an urgent need in this field to find essential hypertension susceptibility genes, and to develop methods, kits, and related therapeutic drugs for detecting essential hypertension.

Contents of the invention

The object of the present invention is to provide a method and detection kit for diagnosing (especially early diagnosis) hypertension.

Another object of the present invention is to provide a new method for treating hypertension.

Another object of the present invention is to provide a pharmaceutical composition for treating hypertension.

In a first aspect of the present invention, a method of diagnosing susceptibility to hypertension in an individual is provided, comprising the steps of:

detecting the individual's ACE gene, transcript and/or protein, and comparing it with normal ACE gene, transcript and/or protein,

A difference indicates that the individual is more likely to have high blood pressure than the normal population.

In another preferred example, the method detects the gene or transcript of ACE, and compares the difference with the normal ACE nucleotide sequence.

In another preferred example, the difference is the following single nucleotide polymorphism:

5525 bits C→G;

Wherein, the nucleotide position numbers are based on SEQ ID NO:1.

In a second aspect of the present invention, there is provided a method for detecting whether there is a single nucleotide polymorphism of angiotensin I converting enzyme gene ACE in a sample, comprising the steps of:

(a) amplifying the ACE gene of the sample with ACE gene-specific primers to obtain an amplified product; and

(b) Detect whether the following single nucleotide polymorphisms exist in the amplification product:

5525 bits C→G;

Wherein, the nucleotide position numbers are based on SEQ ID NO:1.

In another preferred example, the gene-specific primers have the sequences of SEQ ID NO: 2 and 3.

In another preferred example, the length of the amplified product is 100-3000bp, and contains the 5525th position in SEQ ID NO:1.

In the third aspect of the present invention, a kit for detecting hypertension is provided, which includes primers for specifically amplifying ACE gene or transcripts, preferably, the amplified length of the primers is 100-3000bp, And contain the amplified product at position 5525 in SEQ ID NO:1.

In another preferred embodiment, the kit also contains reagents selected from the following group:

(a) a probe that binds to the mutation at position 5525 in SEQ ID NO: 1;

(b) Recognition of the mutant restriction enzyme at position 5525 in SEQ ID NO:1.

In another preferred example, the mutation is selected from the following single nucleotide polymorphisms:

5525 bits C→G;

Wherein, the nucleotide position numbers are based on SEQ ID NO:1.

In the fourth aspect of the present invention, a pharmaceutical composition is provided, which contains a safe and effective amount of an ACE protein inhibitor and a pharmaceutically acceptable carrier.

Detailed ways

After intensive and extensive research, the inventors have determined and analyzed the SNPs of a large number of candidate genes. For the first time, it was discovered and proved that some SNPs of the ACE gene are closely related to high blood pressure, and its new function was discovered: the change of ACE will lead to high blood pressure. G) There is a significant difference in the distribution between the case and the control group (P<0.05), so it can be used as a specific SNP for detecting hypertension. The present invention has been accomplished on this basis.

ACE gene

The detailed sequence of angiotensin I converting enzyme gene (angiotensin I converting enzyme, ACE) can be referred to the nucleotide sequence that accession number is AF118569 (can refer to website http://www.ncbi.nlm.nih.gov/), such as Shown in SEQ ID NO: 1.

The angiotensin I-converting enzyme encoded by the angiotensin I-converting enzyme gene (also called "angiotensin-converting enzyme") has the function of promoting the conversion of angiotensin I into the active phosphorylated polypeptide angiotensin II. Angiotensin II is a potent vasopressor and aldosterone-activating polypeptide that controls blood pressure and homeostasis. This gene plays a key role in the renin-angiotensin (RAS) system.

RAS is the English abbreviation of Renin-Angiotensin-System, and its main function is to regulate blood pressure, water, electrolytes and maintain the stability of the internal environment of the human body. RAS exists not only in the circulatory system, but also in the blood vessel wall, heart, central nervous system, kidney and adrenal gland, and participates in the regulation of target organs. Renin in the blood circulation mainly comes from the kidneys, which can convert angiotensinogen mainly from the liver into angiotensin I, and then convert it into angiotensin II (AngII) under the action of angiotensin converting enzyme, and then pass through the tissue It acts on the angiotensin II receptor. Local RAS exists in many tissues and plays an important role in the function regulation of corresponding organs, tissues and cells. Excessive production of tensin II results in high blood pressure in the circulation. In the tissue, it causes structural remodeling. When the tissue structure remodeling of a certain organ develops to a certain extent, the organ cannot perform normal physiological functions, and corresponding clinical symptoms will appear, such as heart myocardial infarction, kidney disease, etc. uremia, stroke in the brain, blindness in the eyes and so on. Therefore, angiotensin II is not only the cause of high blood pressure, but also the direct cause of heart, brain, kidney and other organ damage.

A number of experiments have been performed focusing on the relevance of the presence or absence of a 287bp Alu repeat within this gene to cardiovascular pathophysiology. In 2002, Procopciuc et al. discovered a SNP, M235T, on exon 2 of the ACE gene (Procopciuc et al. J Cell Mol Med. 2002 Apr-Jun; 6(2): 245-50). This SNP causes a shift from methionine to threonine. In 38 essential hypertensive patients and 21 normotensive controls, they found that M235T was 81.57% in the patient population and 66.66% in the normal controls. Because there is a frequency difference between M235T in the patient population and the normal population, it shows that the ACE gene is a candidate gene for hypertension.

Animal experiments have shown that mice with ACE gene deletion have hypotension, renal dysfunction, female infertility and other phenotypes (Ramaraj et al. J. Clin. Invest. 102: 371-378, 1998).

The inventors sequenced almost the entire region of the ACE gene and found many SNPs, most of which were not associated with susceptibility to hypertension. However, association studies showed that position 5525 C→G was associated with susceptibility to hypertension Very high SNP. The SNP is located in the No. 4 intron of ACE, suggesting that it affects the post-transcriptional splicing process of ACE, which in turn affects the activity of ACE, and ultimately leads to a significantly higher susceptibility to hypertension in carriers than in normal populations.

Based on the new discovery of the present invention, the ACE protein or polypeptide has many new uses. These uses include (but are not limited to): screening substances that promote the function of ACE protein, such as antibodies, polypeptides or other ligands. Screening the polypeptide library with the expressed recombinant human ACE protein can be used to find therapeutically valuable polypeptide molecules that can stimulate the function of the human ACE protein.

On the other hand, the present invention also includes polyclonal antibodies and monoclonal antibodies, especially monoclonal antibodies, specific to human ACE DNA or polypeptides encoded by fragments thereof. Here, "specificity" means that the antibody can bind to human ACE gene product or fragment. Preferably, it refers to those antibodies that can bind to human ACE gene products or fragments but do not recognize and bind to other irrelevant antigen molecules. Antibodies in the present invention include those molecules capable of binding and inhibiting human ACE protein, as well as those antibodies that do not affect the function of human ACE protein.

The present invention includes not only complete monoclonal or polyclonal antibodies, but also immunologically active antibody fragments, such as Fab' or (Fab) ₂ fragments; antibody heavy chains; antibody light chains; genetically engineered single-chain Fv molecules; or chimeric antibodies.

Antibodies of the present invention can be prepared by various techniques known to those skilled in the art. For example, purified human ACE gene products, or antigenic fragments thereof, can be administered to animals to induce polyclonal antibody production. Similarly, cells expressing human ACE protein or antigenic fragments thereof can be used to immunize animals to produce antibodies. Various adjuvants can be used to enhance the immune response, including but not limited to Freund's adjuvant and the like.

Antibodies of the invention may also be monoclonal antibodies. Such monoclonal antibodies can be prepared using hybridoma technology. The antibodies of the present invention include antibodies capable of blocking the function of human ACE protein and antibodies that do not affect the function of human ACE protein. Various antibodies of the present invention can be obtained by conventional immunization techniques using fragments or functional regions of human ACE gene products. These fragments or functional regions can be prepared using recombinant methods or synthesized using a polypeptide synthesizer. Antibodies that bind to unmodified forms of human ACE gene products can be produced by immunizing animals with gene products produced in prokaryotic cells (e.g., E. coli); antibodies that bind to post-translationally modified forms (such as glycosylated or phosphorylated Proteins or polypeptides), which can be obtained by immunizing animals with gene products produced in eukaryotic cells (such as yeast or insect cells).

Antibodies against human ACE protein can be used in immunohistochemical techniques to detect the amount and/or mutation of human ACE protein in biopsy specimens. A preferred anti-ACE antibody is an antibody that does not recognize normal ACE but recognizes mutant ACE, or an antibody that recognizes normal ACE but not mutant ACE. Using these antibodies, the detection of hypertension susceptibility at the protein level can be conveniently performed.

Using the ACE protein of the present invention, substances that interact with the ACE protein, such as inhibitors, agonists or antagonists, can be screened out through various conventional screening methods.

When the ACE protein of the present invention and its antibody, inhibitor, agonist, antagonist, etc. are administered (administered) therapeutically, various effects can be provided. Generally, these materials can be formulated in a non-toxic, inert and pharmaceutically acceptable aqueous carrier medium, wherein the pH is usually about 5-8, preferably about 6-8, although the pH value can be changed according to the Depending on the nature of the substance formulated and the condition to be treated. The formulated pharmaceutical composition can be administered by conventional routes, including (but not limited to): intramuscular, intravenous, or subcutaneous administration.

Normal ACE inhibitors can be used directly in the treatment of diseases, for example, in the treatment of hypertension. When using the ACE protein inhibitor of the present invention, other drugs for treating hypertension can also be used at the same time.

The present invention also provides a pharmaceutical composition, which contains a safe and effective amount of the ACE protein inhibitor of the present invention and a pharmaceutically acceptable carrier or excipient. Such carriers include, but are not limited to: saline, buffer, dextrose, water, glycerol, ethanol, and combinations thereof. The pharmaceutical formulation should match the mode of administration. The pharmaceutical composition of the present invention can be prepared in the form of injection, for example, by conventional methods using physiological saline or aqueous solution containing glucose and other adjuvants. Pharmaceutical compositions such as tablets and capsules can be prepared by conventional methods. Pharmaceutical compositions such as injections, solutions, tablets and capsules are preferably manufactured under sterile conditions. The active ingredient is administered in a therapeutically effective amount, for example about 0.1 microgram/kg body weight to about 10 mg/kg body weight per day. In addition, the polypeptides of the invention can also be used with other therapeutic agents.

When using the pharmaceutical composition, the safe and effective amount of ACE protein or its antagonist, agonist is administered to the mammal, wherein the safe and effective amount is usually at least about 0.1 microgram/kg body weight, and in most cases no more than about 10 mg/kg body weight, preferably the dosage is about 0.1 μg/kg body weight to about 100 μg/kg body weight. Of course, factors such as the route of administration and the health status of the patient should also be considered for the specific dosage, which are within the skill of skilled physicians.

Polynucleotides of human ACE protein can also be used for various therapeutic purposes. Gene therapy technology can be used to treat abnormalities in cell proliferation, development or metabolism due to non-expression of ACE protein or expression of abnormal/inactive ACE protein. The method for constructing a recombinant viral vector carrying the ACE gene can be found in existing literature (Sambrook, et al.). In addition, the recombinant human ACE gene can be packaged into liposomes and then transferred into cells.

The methods for introducing polynucleotides into tissues or cells include: directly injecting polynucleotides into tissues in the body; or first introducing polynucleotides into cells in vitro through vectors (such as viruses, phages, or plasmids, etc.) , and then transplant the cells into the body, etc.

The invention also relates to a diagnostic test method for quantitative and localized detection of human ACE protein level. These assays are well known in the art and include ELISA and the like.

A method for detecting whether there is an ACE protein in a sample is to use an ACE protein-specific antibody for detection, which includes: contacting the sample with an ACE protein-specific antibody; observing whether an antibody complex is formed, which means that the antibody complex is formed ACE protein is present in the sample.

The polynucleotide of ACE protein can be used for the diagnosis and treatment of ACE protein-related diseases. In terms of diagnosis, the polynucleotide of ACE protein can be used to detect the expression of ACE protein or the abnormal expression of ACE protein in disease state. For example, the ACE DNA sequence can be used for hybridization of biopsy specimens to determine the abnormal expression of ACE protein. Hybridization techniques include Southern blotting, Northern blotting, in situ hybridization, and the like. These technical methods are all open and mature technologies, and relevant kits are available from commercial sources. Part or all of the polynucleotides of the present invention can be immobilized as probes on microarrays or DNA chips (also known as "gene chips") for analysis of differential expression of genes in tissues and gene diagnosis. RNA-polymerase chain reaction (RT-PCR) in vitro amplification with ACE protein-specific primers can also detect ACE protein transcripts.

Detection of mutations in the ACE gene can also be used to diagnose high blood pressure. Detection can be against cDNA or genomic DNA. The forms of ACE protein mutations include point mutations, translocations, deletions, recombinations, and any other abnormalities compared with the normal wild-type ACE DNA sequence. Mutations can be detected using established techniques such as Southern blotting, DNA sequence analysis, PCR and in situ hybridization. In addition, mutations may affect protein expression, so Northern blotting and Western blotting can be used to indirectly determine whether a gene has a mutation.

The most convenient method for detecting the SNP of the present invention is to amplify the ACE gene of the sample with ACE gene-specific primers to obtain the amplified product; then detect whether there is the following single nucleotide polymorphism in the amplified product: 5525 position C → G, wherein the nucleotide position numbering is based on SEQ ID NO:1.

It should be understood that after the present invention first reveals the correlation between the SNP of the ACE gene and hypertension, those skilled in the art can easily design an amplification product that can specifically amplify the position of the SNP, and then through methods such as sequencing Determine if there are 5525 bits C→G. Usually, the length of the primer is 15-50bp, preferably 20-30bp. Although it is preferred that the primer is completely complementary to the template sequence, those skilled in the art know that it can also be specifically amplified (that is, only amplify the desired fragment). Kits containing these primers and methods using these primers are within the scope of the present invention, as long as the amplified product amplified by the primers contains the corresponding position of the SNP of the present invention. A preferred primer pair has the sequences of SEQ ID NO: 2 and 3.

Although the length of the amplified product is not particularly limited, generally the length of the amplified product is 100-3000 bp, preferably 150-2000 bp, more preferably 200-1000 bp. These amplified products should all contain the 5525th position in SEQ ID NO:1.

Because the SNP of the present invention has a very high correlation with hypertension, it can not only be used for early and more accurate diagnosis of essential hypertension, but also can take precautions to make the carrier take reasonable preventive measures before the onset of the disease (such as in the Therefore, it has extremely important application value and social benefits.

Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental method that does not indicate specific conditions in the following examples, usually according to conventional conditions such as Sambrook et al., molecular cloning: the conditions described in the laboratory manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the manufacturer's instructions suggested conditions.

Example 1

1.1 Research object

Since blood pressure is a quantitative trait that is affected by multiple genes and is affected by various environmental factors, it is necessary to consider the issue of genetic heterogeneity when analyzing related genes. Selecting isolated populations with a relatively single genetic background as materials for the study of polygenic diseases will help reduce the impact of genetic heterogeneity. reduce. Selecting such a group, whether it is used for linkage disequilibrium analysis or selecting families for family analysis, will help to improve the detection sensitivity of hypertension susceptibility loci. Therefore, in this embodiment, the isolated population is selected as the research object (the isolated population has the advantages of relative consistency of genetic background and relatively small number of founders).

On the basis of informed consent, 324 individuals of Han nationality over the age of 50 were randomly collected from Xiangchang Town, Dabie Mountain, Yuexi County, Anhui Province. 80% of individuals have only 6 surnames, the number of founders should be relatively small, and individuals of the same gender come from the same founder. The selected hypertensive samples require that the systolic blood pressure is not lower than 140mmHg, and the diastolic blood pressure is not lower than 90mmHg, and the blood pressure is measured twice to take the average value.

11% of them are located at the top of the blood pressure distribution (37 individuals; SBP>178mmHg) and 7% are located at the bottom of the blood pressure distribution (22 individuals; SBP<104mmHg), a total of 59 individuals were selected by direct sequencing method Perform SNP detection.

1.2 Experimental methods and results

1.2.1 DNA extraction

DNA was extracted from human blood by the conventional phenol-chloroform method, and the concentration was corrected to 20ng/ul for conventional PCR amplification.

1.2.2 Design of PCR and sequencing primers

According to the genome sequence of ACE in GenBank, the following primers were designed and synthesized. The specific primers are shown in Table 1 below.

Table 1 Primer sequence list Primer name Sequence (5'-3') SEQ ID NO: sense primer gcctgtgtctcagatctcacc 2 antisense primer cagaggcaaagaggagcatc 3

1.2.3 PCR amplification of ACE gene

Using the extracted DNA as a template, PCR amplification was performed on a GeneAmp 9700 PCR instrument with the Touchdown program using Taq enzyme. The reaction conditions are: pre-denaturation at 94°C for 2 minutes, denaturation at 94°C for 30 seconds, annealing at 63°C for 40 seconds, extension at 72°C for 40 seconds, a total of 10 cycles, and the annealing temperature decreases by 0.5°C for each cycle; after denaturation at 94°C for 30 seconds, Anneal at 58°C for 40 seconds, extend at 72°C for 40 seconds, a total of 30 cycles; finally extend at 72°C for 7 minutes. PCR amplification products were verified by agarose gel electrophoresis.

As a result, an amplification product of 395 bp was obtained.

1.2.4 SNP discovery and detection

After the PCR products were purified by Resin resin, the ABI-PRISM ^TM 377 DNA sequencer (appliedbiosystems (ABI)) was used for bidirectional sequencing with fluorescent labeling end termination method, and Polyphred software (University of Washington, USA http://droog. mbt.washington.edu/Polyphred.html) for sequence interpretation and SNP confirmation.

As a result, the following SNP was found to exist: C→G at position 5525.

1.2.5 SNP genotyping and association analysis

SNP genotyping by direct one-way sequencing. That is, typing and correlation analysis were carried out in hypertensive patients and normotensive control groups.

After the allele frequencies are counted, chi-square test is carried out. By comparing the individual data of the highest 11% with the lowest blood pressure of 7%, it is preliminarily determined whether the allele frequencies are linked with the blood pressure level.

It was found that the frequency of the G-type SNP at position 5525 in SEQ ID NO: 1 was 22% in hypertensive individuals and 11% in hypotensive individuals. The frequency difference between the two is 11%, which confirms that there is a correlation between the G/C type SNP at position 5525 in SEQ ID NO: 1 and the occurrence of hypertension.

Example 2

Essential Hypertension Susceptibility Detection Kit

As described in Example 1, the mutation of C→G at position 5525 in SEQ ID NO: 1 is closely related to hypertension. Therefore, based on this mutation, ACE gene-specific primers can be designed to amplify and detect with the patient's DNA as a template.

Prepare a test kit (100 person-times), which contains: name sequence concentration sense primer SEQ ID NO: 2 100pmol antisense primer SEQ ID NO: 3 100pmol PCR reaction solution Containing Taq Enzyme dNTP Magnesium Ion PCR Reaction Buffer

Take 3ml of blood from the male patient to be tested, and use a conventional method (or use a specific kit) to extract DNA from the blood. Dilute the PCR primers in the high blood pressure detection kit to 1μmol/μl, and use the extracted DNA as a template to perform a PCR reaction with the provided primers. After the PCR products were purified, ABI-PRISM ^TM 377 DNA sequencer was used for bidirectional sequencing by fluorescence-labeled end termination method, and Polyphred software was used for sequence interpretation and SNP confirmation.

Alternatively, the chromatographic analysis of the amplified product and the normal control is performed with a denaturing high-performance liquid chromatograph (DHPLC), and the 5525-position C→G can also be detected.

As a result of the test, the susceptibility to hypertension of 5525 test subjects including C→G was higher than that of the normal population.

All documents mentioned in this application are incorporated by reference in this application as if each were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Sequence Listing

<110> Fudan University

  Shanghai Human Genome Research Center

<120>Relationship between angiotensin I converting enzyme gene and essential hypertension

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aacatcaccg cggagaatgc aaggcgccag gtgggcgccc gggcccgggc gggggcgggg 2940

cggggccgcg gcggccaatc acagcacgcg gccggcttgt ggggcgggca ggctggcgcc 3000

cccgacccga accccacccc gaccccggac cctcgccccg acagtcagcc gcggggcccg 3060

agcgccgggc tgcgcgcacg gcctacgctc ccagcatgca cgagttggat ggatgaaggt 3120

ggctgctccc aggccgcgcc cgccttcgcc gaaggtgctg ggcttggctc tggggccccc 3180

gcgctctcgg gcagctgcct tctcacctcc ggacgctgtc gctgtcaccg tcaccgcact 3240

gcactgtcca tccaccctcc actcgcccgg cctcttttct ggtcccaatt tctgctccac 3300

cattcccatg aggcagattc cctccagaag gaggaagcgc ggcttccgca aactaaggtc 3360

tcccgcaggg atctccccag ggcccgggct ctggaagccc ttggccttcc tcccctcccc 3420

cagcaccgtg gcttctcctt tatggcctgc atctaagcag ggtcctacac cctccctgcc 3480

ctcctggtgc ccaataggag gaagcagccc tgctcagcca ggagtttgcg gaggcctggg 3540

gccagaaggc caaggagctg tatgaaccga tctggcagaa cttcacggac ccgcagctgc 3600

gcaggatcat cggagctgtg cgcaccctgg gctctgccaa cctgcccctg gctaagcggc 3660

agcaggtggg ctgagggctg aggcagagct cgggggcggc ctcctagtgc cccatcgtgg 3720

gggtcgggggg agagcagccc atcagggagg gaggaaccct gggatccaca tgggccctga 3780

cagaagggaa agcccaggta agcacagaat ggctttctga gcattgattt ttcttggaga 3840

tggggtgggg agttactttc tgttaaagga agcattctgg agtaggaagc caaattcaaa 3900

tacacttctc cctaggctgg tttatgagct tctttggaag agttgagaag ggctgggcgt 3960

ggtggctcaa gcctgtaatc ccagcacttt gggaggctga ggtgggcgca tcgcttgagc 4020

ccaggagttc aagaccagcc tggccaacat ggcaaaacct cgtctctaca aaaaaaaaat 4080

agctgggctt ggtggtgcgt gcacctacag tcccagctac tcttgaaact gagggggaag 4140

gatcacctga gcccaggagg tcaaggctac agtgagctgt gattgcacta ctgcacccca 4200

gcctgcgtga cagagtgaga cctcccccca aaaaaaagag agagagaaaa ggttgagaaa 4260

gactgggaag tcaccaaagc cagagaatgg gagggatctg ccctcactgc agggtggtgc 4320

caagctggga cttgacccag accctgactt tcaggactcc tgtcccccac tccacaggct 4380

gcctccactg gcaggggact cagaagtgat ccggtcacac taagtgacac ttagtgatca 4440

gaagtgcccc ggtgccactg agtggccttg tccaagctac atccactctg tgggctcctc 4500

cttgtagcag cgaggggagg gcagatgtcc caggggctgg tcactggagc attcctcccc 4560

tctgactccc cagtacaacg ccctgctaag caacatgagc aggatctact ccaccgccaa 4620

ggtctgcctc cccaacaaga ctgccacctg ctggtccctg gacccaggta cggcccttgc 4680

agctcccctc tcggcggtgc cctagtgttc ccacattgcc ctgctgcact ccggaccatg 4740

cagttgtgta gggtctgtgg aagacagcagg taaacccaaa ggtgttgccc tccaactggg 4800

gctggacggt gcagataccc ccacgccctg cttctcttgg caagtggact tccggaatct 4860

ccagctgcag cccccacttc tgtgtgtacc tcggcctctc ccatcacccc taggccttcc 4920

tcctggctgc ctggtttccc ctttcgtggg tcctctcatg ttccccaaga gccctcaggc 4980

cagggacccc tcgtggcttc cccttaaacc ccgctccagc cccctttatg agcagcttcg 5040

aggaaggcac tccatccaat aggccgctaa gtgtctgtct gggttttggc ctttgggtgt 5100

ccccttggtg tcagccacct taggtggtca tgtctctggg gcaggggccc tgcctgggtg 5160

tttctgtagc tcccagcccc ctcccaccag gcctgtaggt ggcccctgtc tctgggggca 5220

ccgtgatgtt caggaagctg gtgggagcag taaggactgg tgcaggctct ggtgaaggcc 5280

gttgaagact tcaacgtgga ggcctcctca ccgaccctgc ctgcctgtgt ctcagatctc 5340

accaacatcc tggcttcctc gcgaagctac gccatgctcc tgtttgcctg ggagggctgg 5400

cacaacgctg cgggcatccc gctgaaaccg ctgtacgagg atttcactgc cctcagcaat 5460

gaagcctaca agcaggacgg tgagcaggcc tctccctgtc caggaaccac gccaggtgtc 5520

ctctctcagc tgtctcccca gagtcccagc ccagagtcag gcagagcagc tggtatgaca 5580

attccagcag gccctgagtt tcccagaaag tggaggtggg accggcctgc acccagtgtg 5640

cctggacttt gctgctggcc tgccccacgt ggccatcctg ctgtcactcc tggccctgat 5700

gctcctcttt gcctctggga acctccagga tctgtttagc tggctgtagc taattagaaa 5760

ttgtagagtg gcaaccccca agccaatttt ccagctagct gcagatccac gggcctcgag 5820

ccagtggaag agccgactta cagctgagag gctgaggtcc gagcctttgg cctgagctac 5880

atacctcacc cccacgcccc caggcttcac agacacgggg gcctactggc gctcctggta 5940

caactccccc accttcgagg acgatctgga acacctctac caacagctag agcccctcta 6000

cctgaacctc catgccttcg tccgccgcgc actgcatcgc cgatacggag acagatacat 6060

caacctcagg ggacccatcc ctgctcatct gctgggtaag gacctggcct cgcctccaca 6120

tgagtcccac ggaagtgtgg gtcccgaggt aggggtgggg gatgtccagg gtaagggaag 6180

gtgggttgtg accctcacat ctcacatgtg tggggcatca tactgtttgc ttcacatgca 6240

ggagaccatt cgtgttccca ctttacaggt ggggaccctg aggcttaggg tcgtgaggga 6300

cttagtggtc agagagctag gggccaaacc aaaggctctg gccctgggtc cagtggggga 6360

gccatcagcc tagctcatgc ccaaggaaac aagcactgtg gccctgcctc aggattgagt 6420

ggctggggcc tggcacagcc agaaatgaca gtggcagcat cttgcagccc caggacatgt 6480

ggccctcgga ggagtgtggg tgggactgat gtgtgagatt tctggcccta agccaggcct 6540

gcagcccttg agggccccag ggtacaggtg ccggccccag ggtgccactc agcgatgcat 6600

gaagaagcag gcacagccag gcagggagcc aagctgtccc cttccttcct tatctagggag 6660

acatgtgggc ccagagctgg gaaaacatct acgacatggt ggtgcctttc ccagacaagc 6720

ccaacctcga tgtcaccagt actatgctgc agcaggtaag ctctgggctc aagcctgggg 6780

tggtgggggt cgggggtggg gcgcaaaaaa agggagtcac agatggggcac aggggcggga 6840

aggtttcggg tactgagcag cagcctggtg tgtctgtagg agcagtgagc tggggtcggc 6900

cccctcagtg aggtgccagc tcctccctcc aggctccaca gtggcaggat gagagcaaca 6960

acgcactttc actcatctgc tgtgggagtg agggccctgc ctctgggaat ggtggccaca 7020

gagcagagaa gctttcatgc acagggagtt gacccgagat ggggaccccca gccctgtccc 7080

caggccagcc agagtgggct ccccctgacc tggctccaca cccctcctcc agggctggaa 7140

cgccacgcac atgttccggg tggcagagga gttcttcacc tccctggagc tctcccccat 7200

gcctcccgag ttctgggaag ggtcgatgct ggagaagccg gccgacgggc gggaagtggt 7260

gtgccacgcc tcggcttggg acttctacaa caggaaagac ttcaggttca gacatgggaa 7320

gagcacgttc tggggttccc cggttctggg gcccggggaa aggcaggcag cccaggcgca 7380

gggaagctgg ttcccaggcc tgcctctacc ctaccccagc actggttgga ggctgggtct 7440

gttccagggc taggggggtat aggaggccta ttagtccacc ttctctggca gctttgacaa 7500

atagtcactt ctataccttg gaatggagga agaaggccca agtggtggtg agccagggca 7560

gggtaaagaa tttgcttgtt tctgccaggc acggtggtca cacctgtaat cccagcactt 7620

tgggaggtca aggcgggtgg atcacctgag gccaggagtt cgagaccagc ctggccaaca 7680

tggcgaaacc ccgtctctac taaaaataca aaaataaatt agccagggggt gatggcggggc 7740

gcctgtaatc ccagctactc aggaggctga ggcaggagaa tctcttgaac ccgggaggcg 7800

gaggttgcag tgagctgaga ttgtgccact gcaggccagc ctgtgcaaaa gagtgagacg 7860

ctgtctcaaa aaaaaaaaga aaaaaagaag ttacttgttt ctactgcggc ttcatgcccc 7920

agggcagctc cctcctcatt cctgtctttc aggtgccaat ctgccctgtg ccctggccct 7980

gccctgttct gtccatccgt cactctcacc ctcgccctct ctacgcccca ggatcaagca 8040

gtgcacacgg gtcacgatgg accagctctc cacagtgcac catgagatgg gccatataca 8100

gtactacctg cagtacaagg atctgcccgt ctccctgcgt cggggggcca accccggctt 8160

ccatgaggcc attggggacg tgctggcgct ctcggtctcc actcctgaac atctgcacaa 8220

aatcggcctg ctggaccgtg tcaccaatga cacgggtatg ggagggctga gaggccccca 8280

cccagcctca cctaaacccc gctccacccc acagcaggac ctcacttgcc ccactcagct 8340

ctgcccttct ttctgcctcc cggccccagg tcaggcaggg ttcgggatcc tcctagagcc 8400

tcacggtgca cactgcgccc agctcagcac acctgggggt cctcttccaa gcagggccca 8460

gggtctcgag ggccagccat accttctctg catctccctg gcctcacttt ctgctgcccc 8520

gccagcccac actcttaggg gaccctcttc tccctctgac ctcttccctc tcctttcatc 8580

tcatctccca acagaaagtg acatcaatta cttgctaaaa atggcactgg aaaaaattgc 8640

cttcctgccc tttggctact tggtggacca gtggcgctgg ggggtcttta gtgggcgtac 8700

ccccccttcc cgctacaact tcgactggtg gtatcttcgg tgagaggagg gatagaaaag 8760

ccttcgcccc agctagccct ccccagcctc ctggacagcc aggcgcctcc tgccccagcc 8820

agttctagcc tctcctctct aatgatgtcc cccgctgtga cccaccgcct tctcctttcc 8880

tgcctgaaac tccctcttcc aggaagtctt ccccagttcc tcaggatggg gaagggttgc 8940

cgggtggaaa tgccttttct acaaaagcta aatccatctg tttgcaacct ctaggcccta 9000

agacaattta accatccttt tccagaacca agtatcaggg gatctgtcct cctgttaccc 9060

gaaacgaaac ccactttgat gctggagcta agtttcatgt tccaaatgtg acaccataca 9120

tcaggtatta gcgcccccac cccacccacc cccagtactg tcacaccctc aatccacttc 9180

tcctcctgtg atcctagctg cctcatcccc agggcttgtc cccatgctcc tccagacctc 9240

aaaggcctgg agttagagtg gcccactctc ctgagcctgt cttgggtctc ccttctcccc 9300

caagatagct tctggtccag cctctgccct gcaggaagct ggatggtgcc tgggtaagga 9360

accccctgttc ctggcccccc atgatcttcc ctgactccca ccctgtgcct gcaggtactt 9420

tgtgagtttt gtcctgcagt tccagttcca tgaagccctg tgcaaggagg caggctatga 9480

gggcccactg caccagtgtg acatctaccg gtccaccaag gcaggggcca agctccggtg 9540

tgtggtggga agccggggga agtgggaggc agagaggagc ggctggcaaa gggtgtggca 9600

ggaggtgtct ggctgctctg atggggtggg gggcaccaac cacagagctg gactgatgtg 9660

gatgcctgtc tcctcgctat gtcatcaaat atttatgag tgggccttct ggctggcatg 9720

gggcgacaca aatgccccct gccaccatca gagagatccc aggccccagg gtcttattgc 9780

cacagtttct gcagtccatt ggggggcgga agtggccagg ggcatgtggg ccggggtcca 9840

ggagcagact ccagcctgag tcccctgtgc ccatggtacc cactctgccc accaggaagg 9900

tgctgcaggc tggctcctcc aggccctggc aggaggtgct gaaggacatg gtcggcttag 9960

atgccctgga tgcccagccg ctgctcaagt acttccagcc agtcacccag tggctgcagg 10020

agcagaacca gcagaacggc gaggtcctgg gctggcccga gtaccagtgg cacccgccgt 10080

tgcctgacaa ctacccggag ggcataggta aagccctgag tgaggatggt gtggggctaa 10140

ggtgggtcct caactctggg cttggcccag gccccaggtt cctggtcagc tcctaccagc 10200

tgagccctgg taccctgtcc tggagggcca ggcagccccc caagctcatc agcagggcct 10260

gcgagtgggg acaggcatgt ctttccccca gcatcctaga gagggtgtgc tcagacctga 10320

gggccctcc ccttccagag gaagccagac acaaggctct gtgaggtcac actgcgggct 10380

ccgctcttat tggccagggg acggtagctg caggactctg ctctcctgcg gccatgggcc 10440

agggttgggc tactgcagga cttcccagcc tcctcttcct gctgctctgc tacgggcacc 10500

ctctgctggt ccccagccag gaggcatccc aacaggtgac agtcacccat gggacaagca 10560

gccaggcaac aaccagcagc cagacaacca cccaccaggc gacggcccac cagacatcag 10620

cccagagccc aagtgggacc atgcaggggga ggggcagggt gccagggggtg ggagaggcgg 10680

ggccgggtag ggacagggca gggtacaagg gagtgcgaga gggataatgg cttctggtga 10740

gaccacaaac ctggagaggg gaggcagagg tttgtctgtt tccctgcact ctgtcccaca 10800

gacctggtga ctgatgaggc tgaggccagc aagtttgtgg aggaatatga ccggacatcc 10860

caggtggtgt ggaacgagta tgccgaggcc aactggaact acaaccacaa catcaccaca 10920

gagaccagca agattctggt gggagccacc tccccacccc caaacctgag catgtgcata 10980

cacacagaga tgctgtcccg ctcaccacac agtggggctg ccaccacatt ttaaattgaa 11040

tattaaaac aatactcaat ttcgggccgg gcgcggtggc tcacgcctgt aatcccagca 11100

ctttgggagg gggaggcggg cggatcacga ggtcagatca agaccatcct ggctaacacg 11160

gtgaaacccc atctctagta aaaatacaaa aattagccgg gtgtggtggc gagcacctgt 11220

agtcccagta ctcaggaggc tgaggcagga gaatggcatg aacccgggag gcagagcttg 11280

cagtgagccg agatggcacc actgcactcc agcctgggcg acagagcgag actccatcaa 11340

aaaaaaaaaa aaaaaaaact caatttcaga ttttgatgaa catttactca atgcctgagc 11400

aattcttctt tccttaaaaa tcagtctctg ggaggcctag gtgggaggat cacttgaagc 11460

caggagttgg agactagcct gggcaacata gcaagatccc atctctattc aaacaaacaa 11520

ataaacaaaa atcaatctct agtaacagaa taatttgtac ataaataagt ggtgctcaag 11580

tcgtttttta aaagattgaa agcctctgtt tgtctcctct acaaaagggg ctacacttcc 11640

tctttaccct cattccctgc ctatttggct gagcacaaat tatgccactg agccaacacac 11700

tgttactgtt ccttggcact ttgatctgtt gcctcatctt tttctcaaca gccttgcaaa 11760

attggtgagc ttaattcccat tttacagatg ggatttgata ttaactctga ggttcagaaa 11820

ggccacagag ctaataccaa gctggctcct tcctaagggc ctttaagaca cttgggggtc 11880

ttctcttctc tgcccctgcc tggatatgtg ttgcttgacc gcaggcatcc agggagggtg 11940

agtactgcat ccaggacgtt atcagcgtcc agcttgcaga gagtcttata ggcaaaggtt 12000

gcaacttaat tccactgccc cctcaccacc acctccagcc ctcagctccc acttggggcc 12060

tcccgctcag aggctgctct ggagctcctg ggccctgtga caccatcccc ctgtgccctc 12120

agctgcagaa gaacatgcaa atagccaacc acaccctgaa gtacggcacc caggccagga 12180

agtttgatgt gaaccagttg cagaacacca ctatcaagcg gatcataaag aaggttcagg 12240

acctagaacg ggcagcactg cctgcccagg agctggagga ggtgtgtggc tcgcaaggta 12300

cagggagggg ggaatcctgg ggcagtgagc ccaacacagg gtctggcctg gccttcacgc 12360

tgcttcctct tcctcgttgt atcaagtcat ggcatctgcc atgcgatgtg cacctcagaa 12420

ctgctgagag ggcagcgctc cccagctccc tggctcccca cctgccagcc catggggcct 12480

gggggtagtg caggccccag agagaccaag tgcaaaggag tacagctcat tgcctctcct 12540

tcctcctgca gtacaacaag atcctgttgg atatggaaac cacctacagc gtggccactg 12600

tgtgccaccc gaatggcagc tgcctgcagc tcgagccagg tgagagctca tgtgcaggct 12660

gagtgagagg cgagggctgg gactggcatg gggcccgggg gtgctgggtg agagcacaga 12720

gttgggttcc cctcgctctt ggggtcagcg tgcccaggaa atgccctttc ttgttttcca 12780

cgaggggggc ttctctgccc cactgagagc cggcacctac ttcataccat gccccgatca 12840

gctgcccctc cctcagaacc gccctctgct taagggtgtc cactctctcc tgtcctctct 12900

gcatgccgcc cctcagagca gcgggatctc aaagttatat ttcatgggct tggactccaa 12960

atggggggaa ctcggggaca ctagctcccc ccggcctcct ttcgtgaccc tgcccttgac 13020

ttcctcacct tctctgtctt tcctgagccc ctctcccagc atgtgactga taaggaaatt 13080

gagtcacaca gcccctgaaa gcgccagact agaacctgag cctctgattc ctctcacttc 13140

cctcacctac cctgccactt cctactggat agaagtagac agctcttgac tgtcctcttt 13200

tctccccact ggctggtcct tcttacccca gcccgtttga aagagctcac ccccgacaca 13260

aggacccgca cacagatacc tcccagctcc ctctcaaccc accctttcca gggttggaga 13320

acttgaggca taaacttgct tccatgagga atctccaccc agaaatgggt ctttctggcc 13380

cccagcccag ctcccacatt agaacaatga caaatagaag gggaaatgga aaataaacag 13440

gagaaacggt tttcccagga cagggtttgg cctacaagtt gtggatgtgg gtacccatgc 13500

caagtgtgag gggaggctgg ccgggtgtgg tggctcatgc ctctaatccc agcactttgg 13560

gaggccaagg tgagtagatc acttgaggcc gggagtttga gaccagcctg gccaacatgg 13620

tgaaacccca tctgtactaa aaatacaaaa gttagctggg cgtggtggta gatgcctgta 13680

gtcccagcta cttgggaggc tgaggcatga gaatcgcttg agcccagcca gggcaataca 13740

gcaagacccc gtctctacaa ataaaataca aaaaattagt tggatgtggt ggtgcatgcc 13800

tgtagtccta gctgctaggg aggctgagat ggaaggattg cttgagcctg ggaggtcaag 13860

gctgcagtga gccgagatgg cgccactgca ctccagcctg ggcaacagag tgagaccctg 13920

tctcagaaag aaaaaaaaaa aaaaaggaga ggagagagac tcaagcacgc ccctcacagg 13980

actgctgagg ccctgcaggt gtctgcagca tgtgccccag gccggggact ctgtaagcca 14040

ctgctggaga ccactcccat cctttctccc atttctctag acctgctgcc tatacagtca 14100

cttttttttt ttttttgaga cggagtctcg ctctgtcgcc caggctggag tgcagtggcg 14160

ggatctcggc tcactgcaag ctccgcctcc cgggttcacg ccattctcct gcctcagcct 14220

cccaagtagc tgggaccaca ggcgcccgcc actacgcccg gctaattttt tgtattttta 14280

gtagagacgg ggtttcaccg ttttagccgg gatggtctcg atctcctgac ctcgtgatcc 14340

gcccgcctcg gcctcccaaa gtgctgggat tacaggcgtg atacagtcac ttttatgtgg 14400

tttcgccaat tttattccag ctctgaaatt ctctgagctc cccttacaag cagaggtgag 14460

ctaagggctg gagctcaagg cattcaaacc cctaccagat ctgacgaatg tgatggccac 14520

atcccggaaa tatgaagacc tgttatgggc atgggagggc tggcgagaca aggcggggag 14580

agccatcctc cagttttacc cgaaatacgt ggaactcatc aaccaggctg cccggctcaa 14640

tggtgagtcc ctgctgccaa catcactggc acttgggtcc cttcattttc ctcaaagagg 14700

tgctgtgaaa ccccaagcct aggaaaaggt agatccctgg aggaggcagg taatgtggtg 14760

ttgggagagc ctggctgtgt cccctctgta ggctatgtag atgcaggggga ctcgtggagg 14820

tctatgtacg agacaccatc cctggagcaa gacctggagc ggctcttcca ggagctgcag 14880

ccactctacc tcaacctgca tgcctacgtg cgccgggccc tgcaccgtca ctacggggcc 14940

cagcacatca acctggaggg gcccattcct gctcacctgc tgggtaaggg cacatgtcgg 15000

gccttgagga gggtaaagac ggaccacagt gtgagtgagg gttgggacag ggctgactag 15060

agggtaggga gcaggctggg gactgagaga ctccagccct gtgggggatg gttgcccagg 15120

ctggaggggg gtgggcgctg ggagtgggga gccccccact tgcatctggt gccacattca 15180

ctgcagatct atgtcgggca agtcaccatg gatgggggaa gaagttaata atcttgtcca 15240

ggagaccacg gcacccatca caacattgtg tgatcttaga gggcgaggaa gaggctgtga 15300

gtgggagctg gggaggcttt gccaagaggt ggcctgtgag cagggcctcg gaagatgaca 15360

gggtttgaca gatgggaagt gggggatgag aggacagacg cagtgttcag gccaagggaa 15420

ctggaacaaa gaagaacctg agaatgtaaa tctacttcaa ccctggaccc tcctttgcca 15480

agggctgcaa tctcagatgc cctgaatgtg tgaagtaggc ggtgaggaca gtaagggatg 15540

gtagggagta aggcaaagca gaggctactg gttctctgtc cctgatgggc tgttaggaac 15600

actttcctgg agcagagaga ccagacaggc cctcagacca tttagaaact ataagggagg 15660

ccccagagga cggcctggct gtgggtctag ctcccacaca ggctgggagt ccagccctct 15720

tcagcccctc tctggtgaga ccaaagaaca tctggtgatg tcacagtgga cgtcagttca 15780

ccaactggga gacacaggcc ccgggaagaa aagcaacatg cccagcgtgg cctgggagct 15840

ggggcagagc tggccttaga actcagcccc tgacaattgg taaaagggga aaggggagca 15900

acctaacact gatgcgctct ctgtctctct ctctggctct ctccctggct ctctccctct 15960

tctctctcat gttctctcca tcactcatcg ctctaaccct ctctcactgg tttgcacttt 16020

agacttcatc tgatgtcagc cgaagcttca cctcacttgg ctaggaaaga gccttgagtc 16080

caaatctgtt tctgagcctt ccattcatcc tgagtttctt ccttttcctc tgtcgtggag 16140

aactaggctc ttttcttaca ctaaactcag aggcatcagc ctctcctgaa ggagacggct 16200

ggttcctgtc agagttgctg agctgcagac accgacctca ggtggtgcgg agggggacatg 16260

gcagagtggc tggtgaagag agcagcctgc cagcctttca atcccagccc tgccacttag 16320

gagccgtggg cccccggcga ggggggcggt cacttaactc tccagcctgt ttcctttact 16380

agccaatggg aatcgtgaca gtacctgggt gcagacagga ttgaaagtga attcacacaa 16440

tgttcttggt gcagagccaa taagaggtgg ccaccggggt gtaggtgttc tggggacctg 16500

taatgtcctc acatgtcagc agttgctagt cacattggtc tccactgctc acggacagtg 16560

aagaccacct ggattccctg ataaccagca aggcccccac ctagagccag gcagtaatga 16620

cctactgggc tggatatgca caccaaagat gatgtgtgcc tcaaagcttg caaacagtag 16680

gtgctcaaga aatgccacta tgattagcag gactgggatc tggagcgctc ttcctgcagg 16740

agggcattga gcctaagtaa catttgtctt tcctctctct gccgtccccc acactcgcct 16800

ccagggaaca tgtgggcgca gacctggtcc aacatctatg acttggtggt gcccttccct 16860

tcagccccct cgatggacac cacagaggct atgctaaagc aggtccgcac cagcccaggg 16920

gcagggaggc cccgccggga tgggagggac cctctgattc aggagttccc tccagtttag 16980

ccctcccccg ggatccccac ggcagcacgc agtctgtccc cggaaccccc agtttgggca 17040

gaactccctc tgcttgcagg gctggacgcc caggaggatg tttaaggagg ctgatgattt 17100

cttcacctcc ctggggctgc tgcccgtgcc tcctgagttc tggaacaagt cgatgctgga 17160

gaagccaacc gacgggcggg aggtggtctg ccacgcctcg gcctgggact tctacaacgg 17220

caaggacttc cggtacatcc agctagggct caggtctcgt tcctgagccc cacgggcaag 17280

ggaaatgaac caagcaaagg gtccactact gtcccccagc tggagccagc agggcaggat 17340

ggggacaggg ccagagtttg ggactgagtg tctagagagg tgttggcttc tggcaggaaa 17400

acccccatccg cctgatgggg acttctgaag cacgcaacag ctctgtcagc ctggccgctg 17460

ggaagtgctc aaggtccccag tcctgggttt gagcatggta ggctgccccg cgtccctcct 17520

tgggagcagc ccctgcatgg agctggcctc tccctggggg cacatgctgt gacacaggga 17580

ggcacacgag gatgttgggt gctctgtaca gatccactct cacccctgac aggctcagaa 17640

gctgccttcc ttggaggatg gcgttttagt tacctattgc tgtgcaacca agcaccccag 17700

agcttagcct tacgaaacaa ccagttgatt ttgcttatga ttttatgtgc caggaattca 17760

ggcagtacac agtggaaatg gcctttctct acagggcccc ctctgttggg ggcagctctc 17820

acagccggga tggctcaatg ggggccatac gtccagagcc ccagttctgg ctgtcggttg 17880

aggtcctcgg tttttcccat gtggcagatg ctggggcaca tgttcccagt ggcctcttgg 17940

ctcacatgtt gggtgcttgg ggtgagatgg ctggaacggc tggaggtggg tcaggcatct 18000

ctccaggcta gctcgggcgc cctcccagcg tggaatctga ggtgggcaga tttacctggc 18060

agccagcatc ccccacagca actactgacg cagccagttc tcaaggctag gtccaaaact 18120

ggcccagagt cacttctgcc atgttttat ggctagaaca agtcacaagt tcacccagat 18180

tcaagagaag agaaaaaagt ccctcccact tgggagaagt ggcaaagacc atctgtcaca 18240

gctgaagaag tgtctcttac aaggagaaca gacacgggga gcctgaaaca aaacccgatg 18300

ggattccctg ggctgtgcag gcccttccag gcatgaggac tcagccacag ggctgagagg 18360

agacaggatc tgggggatga gagcccttgt ggggtcttcc cttttatggg gagtcagagg 18420

agaagctgga tagatcccca gccttgtggc caggatgctg ggcagctctt ccttccccct 18480

ccccgatgag aatgacagaa aaacaggatt cacctgagcc aaaaggcttc cagttagatc 18540

caagagagaa tttcccgcag tttgaattgg tttgctaaac aacaaggaag ggctgggtgc 18600

ggtggctcac acctgtaatc tcagcacttt gggaagccga ggcaggaggt ctgcttgagc 18660

tcaggggttc gagaccatcc tgggcaacat agcgagaccc catttcataa aaaataaata 18720

agtaaatgag aacaaggaag gactgacgag agacggtaga accttctggt ttgggcagct 18780

ctgcagctgc cattcatcct ggccataaga attcttgggg tgaataagtt tgtcgctgtt 18840

gggccgcatg agatgcagaa tcgcccactc tcacccctga cagaaacagt tgtttccttc 18900

agggagcctc catcttggga gataaagcat gtgtacatgg gaacccactg gccacacatt 18960

ctctagaaag tacacaatgt cccagtgcct ctagagcaag cactttgtac agtcagaaag 19020

caacaggtgg tgggggctgg agtcattcag gaaaatggga ggcagaggaa tggcctgaac 19080

ggcccgatgc tagggggcttc tgcccccaga ttccctctta cgcacactca gtggttgccc 19140

ttcccctccc tccccacagt gctgtgtccc ctgcatgctg cagtgctggg gtctgccctg 19200

ggtatagcaa ggcccactgt tcccttatgc ccagggcttc tcactgtcct ctcccaacac 19260

cctctccccc actccactat tcctaggatc aagcagtgca ccaccgtgaa cttggaggac 19320

ctggtggtgg cccaccacga aatgggccac atccagtatt tcatgcagta caaagactta 19380

cctgtggcct tgaggggaggg tgccaaccccc ggcttccatg aggccattgg ggacgtgcta 19440

gccctctcag tgtctacgcc caagcacctg cacagtctca acctgctgag cagtgagggt 19500

ggcagcgacg gtgagagaga agcgggaggc cctggtgggc tgaggaccaa gaaagggtgg 19560

tgagcttggg aggtgggaaa ggggcactta gtggcccatg ggcagaggtg tggggcagag 19620

caatcggaag gaagggagcc accccagacca tcccaggagg caggtcacag ggcccaaaag 19680

gtacagcacc cccaccccctc caccatcaca ggcacaccag ggccaagccg ctaggaccct 19740

gggtctgaca gctgggctcc cttcccttgc agagcatgac atcaactttc tgatgaagat 19800

ggcccttgac aagatcgcct ttatccccctt cagctacctc gtcgatcagt ggcgctggag 19860

ggtatttgat ggaagcatca ccaaggagaa ctataaccag gagtggtgga gcctcaggtt 19920

ctggaacact cccacgggat gcgggctggg ggatctctgc gagtgtctgc atgtgcctgg 19980

gtgtctggat gggccagggt aggggaggtgt gtgtgtgtgtgt gtacactatt gtgtctgtgc 20040

atatgatgtg tgtggtgtaa gtgatgggga aaacggggtg attgtgcaca gaggcccagc 20100

acgcaggaga atggggtgcc cagtatagcc ccaagtgcag ggaccctccc tcaagtcaaa 20160

aatgccacccc ccagcctggt tctccccaaa ctcatcttcc aacatatatt cccactcgac 20220

aggctgaagt accagggcct ctgcccccca gtgcccagga ctcaaggtga ctttgaccca 20280

ggggccaagt tccacattcc ttctagcgtg ccttacatca ggtaacggga aaggcaggag 20340

ggcacattgt gaggggcagt accccacagct ttgtgtttca actgcggcca ctgcccggtc 20400

cacaagctct gtcagtcagg gcagacccgg gggagccggc cgcacggtgc aggtgcctgg 20460

gcccactcac actgccaagg ctgatgggtt ttttcttgaa cattcttttg atgagagtct 20520

gtaccatcca aacagttgaa cacagaaact caacctaata attggctaat ggttaccaga 20580

ccttggttaa gtagttaaca ttaaccacga ctcatggctg gatcatgagc tctgcactgt 20640

tttgttttgc ttttaaaaca agactgtgat tcttttacta ttaattgaaca ttgtctgcga 20700

tacaatttga attgtacctg gaagcccttc tagacactaa aatgtaggat tggagatcgg 20760

ttaaggtggg aggcagggtt gctggggcaa gttacagtca caggctgggg tcagacagaa 20820

ctgggttcaa accccgtctc cattactttg tttcccttgag aaaattcctc aatttctgtg 20880

agcttccatt tcctgacctg tgaaccccat ttcacaggat gcacgatggc taacttctta 20940

gcattctgtc tcatacacag cctcctcagg gaggggtggc caggaccccca ctattcatca 21000

ctctctagtg gaatgtagct gcacactagg tctgcaggtc acatggccac agatgagtgt 21060

gcccaatgca gcccctctcc ttctgtgtgc cccgggagag cacttgctga gggctagcaa 21120

ggctgtttgt gatccggggag gctccctggg aggctggggg ctagagagac ctcaggctgg 21180

agttccaggt gcccccgggc tacaggtagc ccaggccacc cccagagggc tgtggctgcc 21240

tctggccctg gcctcccgtg gttcctggaa gcccagcaag ggcagggccc atgcccacct 21300

tgcctcctgg cacctgggat gatgccagca catcatcaag tgctaataac tgattgtggg 21360

atggatgaag tctgtcccca gagtccagga agagggcatc cctggagcac ctcagatagg 21420

cctgacctag acggtgctcc agagatgaca cttaggacag ggctcccgcc tgcctgctgg 21480

agtggtccct gggttccca gccggcgctg gctctcaccc gggagccagc tggtgtgatg 21540

gctagcttcc cagcttaatg cagacaattc tccaaacagg ggtgggcaaa ggagacttgg 21600

ctgctctaga aaaacattcc ggattctggc cagcagcctt cacaaagcac ttttaggaaa 21660

gaccagggaa ctaggtggta catgcttgca cccagcattc aaagtgagag gcctgtgcca 21720

ctggctcagg acatttaaaa cctcttcaga ctttaagctg gggagaatcc tccagccttg 21780

actggcagat ttctaccagg gaattcgtga tgctttggat aagatcatgt aggactggct 21840

tccctgccca gaccacccta gtagatccac acggcccatt tggccacacc ttgcctctac 21900

ttgcatatac cccagggatg gagacctcac tgcctccaaa gccacctgcc agatctctga 21960

aggctctgcc ctgaccccat ggagcaggcc ctcctgagtt gtgggaggcag ctctgtgggt 22020

gggaggcatc tacacaggca cggctaggaa gaggctcaga caatgctaag agctggggtg 22080

ggggagctca ccctgatagc tgtgggcaga gttggggggc cttggctctg ctgtgcgcat 22140

gtgacttagc acacatcaca tgtgatgtgc agaagggcct ggggcccagt ggcacaaggc 22200

cctcaaccaa ctccgccccg ggccacggcc tcgctctgct ccaggtactt cgtcagcttc 22260

atcatccagt tccagttcca cgaggcactg tgccaggcag ctggccaacac gggccccctg 22320

cacaagtgtg acatctacca gtccaaggag gccgggcagc gcctggcgtg agtgtcctcc 22380

agccctcctt tgtttccatg ctctggcctg cgcccctggg ccttgagggg tctgtccact 22440

ggagcttttg tgggaacact tgccattttg agccgggaac tcccacctgc agcgtgggcc 22500

aggcctgatt gccatctcct taggcacctg gagccctggg gccctgggac aagtttcagc 22560

tgggagtggg tatggagagt ggatgtcagg tgggggcaag aggggccatg tccttctgac 22620

tctgcctccc tgtctcatgc ctccccagga ccgccatgaa gctgggcttc agtaggccgt 22680

ggccggaagc catgcagctg atcacgggcc agcccaacat gagcgcctcg gccatgttga 22740

gctacttcaa gccgctgctg gactggctcc gcacggagaa cgagctgcat ggggagaagc 22800

tgggctggcc gcagtacaac tggacgccga actccggtac cgccaccac cccacctcca 22860

gccttgggtc ttaaccccct ccccaggctg ggcagccatg cggctgacct cggagcctgg 22920

ccctgccccg cacccttgcc ctgccctgcc ctgccctgcc catgctgtct ccttgcttcc 22980

cactcagctc gctcagaagg gcccctccca gacagcggcc gcgtcagctt cctgggcctg 23040

gacctggatg cgcagcaggc ccgcgtgggc cagtggctgc tgctcttcct gggcatcgcc 23100

ctgctggtag ccaccctggg cctcagccag cggctcttca gcatccgcca ccgcagcctc 23160

caccggcact cccacggggcc ccagttcggc tccgaggtgg agctgagaca ctcctgaggt 23220

gacccggctg ggtcggccct gcccaagggc ctcccaccag agactgggat gggaacactg 23280

gtgggcagct gaggacacac cccacacccc agcccaccct gctcctcctg ccctgtccct 23340

gtccccctcc cctcccagtc ctccagacca ccagccgccc cagccccttc tcccagcaca 23400

cggctgcctg acactgagcc ccacctctcc aagtctctct gtgaatacaa ttaaaggtcc 23460

tgccctcccc atctgagtct gtgtccctca cagggaagcc agggacagggg acaggctgct 23520

ttcctgcctc ctggcagtca agtgggtccc gttactaggt ttgttcctcc atcctccttc 23580

aggagccggg gaggatcccc agagctctgc cccagcacct cctggcgctg gcgcctgtct 23640

tccctccagc ccaggcagcc cgccactgtc ctgccaccgc aggcagcccc tgtctggccc 23700

aagcactgac ccacgcggac tctgggaagc agacatcctg ggctgctggc ctcacatttc 23760

cactggcagt ggagcctttc cctgctccac aaatggccag gtccccccag gggaaggctt 23820

ccggctgtta tcggctgcct cagggggcga gtaccttgga gggcctgctt caaggagggt 23880

gccccctgga gggcacacac cagcctagtg cttaccttgg ctcctgcctg taccagctcc 23940

atgactctct gctcgggtga acagccttgg ctctcagaca gccattctaa cactgccagt 24000

gcagaggggc ctcagacgct ggagtgtagc agtggctgca cctgcacagg gattagctgc 24060

cagcagccac 24070

<210>2

<211>21

<212>DNA

<213> Artificial sequence

<220>

<221>misc_feature

<223> Primer

<400>2

gcctgtgtct cagatctcac c 21

<210>3

<211>20

<212>DNA

<213> Artificial sequence

<220>

<221>misc_feature

<223> Primer

<400>3

cagaggcaaa gaggagcatc 20

Claims (10)

1. A method for in vitro detection of whether there is a single nucleotide polymorphism in angiotensin I converting enzyme gene ACE in a sample, characterized in that it comprises steps: (a) amplifying the ACE gene of the sample with ACE gene-specific primers to obtain an amplified product; and (b) Detect whether the following single nucleotide polymorphisms exist in the amplification product: 5525 bits C→G; Wherein, the nucleotide position numbers are based on SEQ ID NO:1. 2. The method of claim 1, wherein the gene-specific primer has the sequences of SEQ ID NO: 2 and 3. 3. The method according to claim 1, wherein the length of the amplified product is 100-3000bp, and contains the 5525th position in SEQ ID NO:1. 4. A test kit for detecting hypertension, characterized in that it includes primers for specifically amplifying ACE genes or transcripts, and the amplified length of the primers is 100-3000bp, and contains SEQ ID NO: 1 The amplification product at position 5525. 5. The kit of claim 4, further comprising reagents selected from the group consisting of: (a) a probe that binds to the mutation at position 5525 in SEQ ID NO: 1; (b) Recognition of the mutant restriction enzyme at position 5525 in SEQ ID NO:1. 6. test kit as claimed in claim 5, is characterized in that, described mutation is following single nucleotide polymorphism: 5525 bits C→G; Wherein, the nucleotide position numbers are based on SEQ ID NO:1. 7. test kit as claimed in claim 4, is characterized in that, described primer has the sequence of SEQ ID NO:2 and 3. 8. A method for diagnosing an individual's susceptibility to hypertension, characterized in that it comprises the steps of: detecting the individual's ACE gene, transcript and/or protein, and comparing it with normal ACE gene, transcript and/or protein, A difference indicates that the individual is more likely to have high blood pressure than the normal population. 9. The method according to claim 8, characterized in that the gene or transcript of ACE is detected, and the difference is compared with the normal ACE nucleotide sequence. 10. The method of claim 9, wherein said difference is the following single nucleotide polymorphism: 5525 bits C→G; Wherein, the nucleotide position numbers are based on SEQ ID NO:1.