KR20040009054A - Gene testing method for finding character of each gene of human - Google Patents

Abstract

PURPOSE: A gene testing method for finding distinctive character of each gene of human is provided, thereby preventing and treating various genetic diseases through gene reconstitution personality. CONSTITUTION: A gene testing method for finding distinctive character of each gene of human comprises the steps of: selecting one primer from primers for testing genetic diseases or personality; amplifying the primer by PCR; subjecting the amplified primer to agarose gel electrophoresis; and determining the distinctive character by verifying the band pattern developed, wherein the gene consisting of the primer for finding genetic diseases comprises dementia gene(apoE), lumbago gene(HLA-B27), corpulence gene(β-ARG), osteoporosis gene(VDR), alcohol resistance gene(ADLH2), diabetes gene(IRSII), lung cancer induction possible gene(CYPIA1), hypertension gene(AGT) and arthritis gene(ER); and the gene consisting of the primer for finding personality comprises physical strength gene(ACE), poisoning gene(DRD2), curiosity gene(DRD2, 4, Intron6) and depression and violence gene(Serotonin transporter).

Description

Gene testing method for finding character of each gene of human}

The present invention relates to a genetic test method that enables the use of various useful information by grasping the characteristics of individual human genetic factors.

In particular, amplification of primers designed according to the nucleotide sequence of a specific genetic factor using a characteristic of a polymerase chain reaction (PCR) to confirm various personal characteristics such as genetic diseases, constitutions and toughness through mutation patterns of the genetic factors The amplified product thus relates to a genetic test method for individual characterization of the human genetic factors provided to determine whether the mutation of the genotype by agarose gel electro-interlocking.

As the genetic engineering advances, the proposed method to amplify specific DNA sequences through the characteristics of polymerase chain reaction and to identify the variation patterns of various genetic factors is used for the examination of diseases of animals and plants as well as humans. It is developing until.

However, in order to perform high-quality diagnostic work using the above method, even if the development of primers using the nucleotide sequences of the relevant genes and the conditions of PCR corresponding to them must be sufficiently presented, they are not satisfied. It resulted in a significant impediment to reproducibility.

Therefore, for the efficient inspection work, the priorities of the above conditions must be taken into consideration.

In order to satisfy the above requirements, the present invention uses the genetic factors of the subject to identify various genetic diseases, as well as personal information such as health and personality such as constitution, physical strength, etc. It provides a genetic test method that allows DNA genes to be identified by PCR and agarose gel electro-linking using primers designed to sequence DNA factors related to disease and toughness. There is a purpose.

In order to achieve the above object, the present invention selects one from each of the primer sequences prepared as genetic factors for genetic disease or toughness test, and then polymerase chain reaction (PCR) so that the presence or absence of genetic mutation of each primer can be confirmed. ) And the size of the product is confirmed by agarose gel electrophoresis so that the band pattern of the genotype can be identified.

At this time, the genetic disease genes include dementia gene (apoE), low back pain gene (HLA-B27), obesity gene (β-ARG), bone covalent electron (VDR), alcohol resistance gene (ADLH2), diabetes gene (IRSII) ), Lung cancer-causing genes (CYPIA1), hypertension genes (AGT) and arthritis genes (ER), etc. will function as targets.In contrast, the genetic factors for testing personality (ACE) and toxic genes (DRD2) ), Curiosity genes (DRD2, 4, Intron 6), and depression and violence gene (Serotonin Transporter), etc. may be proposed as the object.

The genetic test method of the present invention can be reproduced in a good state through the following description.

First, looking at the genetic characteristics of various diseases that can be applied to the present invention,

Dementia is well known as a disease that causes memory impairment and affects both daily life and occupational activities. The dementia gene suggested for the treatment of such disease is apoE, and the test method of the present invention is the base sequence of this gene. Four types (E2 / E3, E2 / E4, E3 / E3, E3 / E4, and E4 / E4) are used to identify people with E4 type who are most likely to have dementia. .

The low back pain gene test is a test for the possibility of ankylosing spondylitis, which is known to be related to low back pain. Such ankylosing spondylitis is known to be caused by the interaction of the human HLA-B27 gene with certain pathogens. The test is expressed in two types, (+) / (-), and the (-) type is known as the genetic type without the possibility of ankylosing spondylitis.

Obesity gene testing, unlike dietary habits and environmental factors, the diet is not very effective when the obesity due to genetic defects, β-3AR genes are recognized obesity genes, respectively, these are TT, TA, AA There are three types, of which TA and AA types are reported as obesity gene types.

Osteoporosis and arthritis testing is a disease of bone that has important functions to support the body and maintain the skeleton. It has been reported that osteoporosis and arthritis are caused by variations in several genetic factors. Representative genes are VDR (Vitamin D Receptor) and apoE, estrgen, collagen, PTH (thyroid) genes are also present.

At this time, the VDR gene exists as BB, Bb and bb types, and the ER genes include PP: xx, PP: XX, pp: XX, and Pp: Xx types, of which bb and PP types have the potential for osteoporosis and arthritis. It is reported to be high.

The alcohol resistance gene test is a test for the ability to resist alcohol. The amount of alcohol resistance is determined by the rate of alcohol degradation depending on the amount of the gene ALD.

There are four types of the gene ALD, among which the modified ALD2 gene, which has the highest activity, is not able to effectively decompose acetaldehyde and thus has low resistance to alcohol, and this alcohol-resistant gene type is G / G (1 / 1), G / A (1/2), A / A (2/2), and G / G (1/1) type is known to have the highest resistance to alcohol.

The Diabetes Genetic Test is a screening test for the development of diabetes. The analysis of skin cells in diabetic patients reveals the protein PC-1 and Mt16189, a gene that makes the protein resistant to the normal activity of the insulin hormone. Check with PCR to determine the likelihood of developing diabetes.

Lung cancer testing is known to have a combination of p53, a type of anti-cancer gene, and the CYP1A1 gene, which regulates the secretion of cytochrome enzymes. As a test for lung cancer, these lung gene promoters can detect the mutation type of this gene by examining the sites of CYP1, CYP2, and GSTM1. In particular, the three types of mutations include CC, 1 with all three mutations. CP with two-part mutations and PP-type with all three mutations were classified as PP type. Among these, CC type was reported to be most likely to cause lung cancer.

Representative genes related to blood pressure gene test and hypertension include ACE (Angiotensin Coverting Enzyme) and Angiotensinogen (ACE), which are also known to be related to eating habits and traditional lifestyles. , gg type genes are present, and GG type has been reported to be associated with high blood pressure.

Next, looking at the human genetic factor,

The fitness gene is made of primers using the base sequence of ACE (Angiotensin Converting Enzyme) gene, which has been known for a long time in heart disease research, and has three types such as II, ID, and DD through the form of PCR. It can be divided into types.

Type II is the strongest of the three types, the ID type is medium, DD type is known to tend to have the lowest physical strength. Compared to the type II, DD type is twice as strong as the force to contract blood vessels It is known that DD type requires stronger heart rate than type II because it requires a lot of nutrients and oxygen when exercising or overworking.

Poisoning gene is a test for predicting attachment and toxicity among the innate personality of subjects by analyzing mutations of dopamine receptor (DRD2A, B) genes, and DRD2 (Dopamin Receptor) The primers were designed using the nucleotide sequences of D2 A and B), and three primers (A1 / A1: B1 / B1, A1 / A2: B1 / B2, A2 / A2: B2 / B2) were used. PCR mutations are used to determine human addiction.

The curiosity gene test is a test for predicting the innate character of the subject, namely, the degree of excitability, impulsivity, and disorder by analyzing the genetic variation of the dopamine receptor. Of these, the 1/1 type is reported to have strong curiosity and impulse.

Depression and Violence Genetic Testing is a test that predicts the possibility of depression and violence among the innate personality of subjects by looking for mutations in serotonin receptor genes.

Therefore, in the present invention, after performing the conditions of the polymerase chain reaction (PCR) suitable for the genetic mutation of each of the primer sequences designed as the genetic disease or the human diagnostic factors for characterization to determine the size of the product Agarose gel electrophoresis confirms the band appearance.

At this time, the extraction of the genetic factors necessary for each genetic disease or toughness is preferably extracted from the hair root of the human body in general, that is, 3-5 strands of hair roots using the sterilized scissors of the human hair. After cutting about 2 cm, it was transferred to a 1.5 ml tube, and again suspended in 200 µl of 5% Chelex, 20 µl of protease-K (20 mg / ml) solution and soaked at 55 ° C. for 1 hour. After boiling, the water is boiled and heated for 10 minutes, and the supernatant is transferred to another tube through centrifugation and applied to each PCAL condition.

Hereinafter, each preferred embodiment of the present invention will be described in more detail.

EXAMPLE 1

Example 1 presents a method according to PCR (PCR) conditions, primer sequences, band patterns and buffer preparation for diagnosing mutations in dementia genes.

Apo E (dementia gene)

※ primer sequence

F: 5'-TCG GCC GCA GGG CGC TGA TGG AC-3 '

R: 5'-CCC AGG CGC TCG CGG ATG GCG C-3 '

※ 5X Buffer Recipe

Mix 10x buffer and 100% glycerol 1: 1 to get 5x buffer.

※ HhaI (Takara) 37 ℃ 2 hour 4% agarose gel

※ enzyme reaction

PCR product 15 ul

Enzyme ul

EXAMPLE 2

Example 2 presents PCR (PCR) conditions, primer sequences and band patterns for diagnosing mutations in low back pain genes.

HLAB-27 (back pain gene)

※ primer sequence

F: 5'-CAGTC TGTGC CTTGG CGTTG C-3 '

R: 5'-GCTAC GTGGA CGACA CGCT-3 '

EXAMPLE 3

Example 3 presents PCR (PCR) conditions, primer sequences, band patterns and enzyme reaction conditions using the obesity gene β-3 Adrenergic Receptor.

beta-3 Adrenergic Receptor

※ primer sequence

F: 5'-CGC CCA ATA CCG CCA ACA C-3 '

R: 5'-CCA CCA GGA GTC CCA TCA CC-3 '

※ Mva I 37 ℃ 2 hour 3% agarose gel

※ enzyme reaction

PCR product 12 ul

enzyme 0.7ul

EXAMPLE 4

Example 4 sets forth PCR (PCR) conditions, primer sequences, band patterns and enzyme reaction conditions for diagnosing mutations in osteoporosis genes.

VDR (osteoporosis gene)

※ primer sequence

F: 5'-TGC TCA TGG CCA TCT GCA TCG T-3 '

R: 5'-CTA ACC AGC GGA AGA GGT CAA G-3 '

※ Bsm I 65 ℃ 2 hour 2% agarose gel

※ enzyme reaction

PCR product 12ul

Enzyme 0.7ul

[Example 5]

Example 5 presents PCR (PCR) conditions, primer sequences, band patterns and enzyme reaction conditions using MT16189 used as a diabetic gene.

MT16189 (diabetic gene)

※ primer sequence

F: 5'-CCA TTA GCA CCC AAA GCT AA-3 '

R: 5'-GTA ATG TGC TAT GTA CGG TA-3 '

※ Mnl I 37 ℃ 2 hour 3% agarose gel

※ enzyme reaction

PCR product 12 ul

Enzyme 0.7ul

EXAMPLE 6

Example 6 sets forth PCR (PCR) conditions, primer sequences, band patterns and enzyme reaction conditions for diagnosing mutations in arthritis genes.

ER (arthritis gene)

※ primer sequence

F: 5'-CTC CTT ACA TTC CAT CTG CTG AA-3 '

R: 5'-CAC CCT GGC GTC GAT TAT CTG A-3 '

※ Pvu II 37 ℃ 2 hour 2% agarose gel

Xba I 37 ° C 2 hour 2% agarose gel

※ enzyme reaction

PCR product 12 ul

Enzyme 0.7ul

EXAMPLE 7

Example 7 sets forth PCR (PCR) conditions, primer sequences and band patterns for diagnosing mutations in fitness genes.

ACE

※ primer sequence

F: 5'-GCCAC TGCTG GAGAC CACTC-3 '

R: 5'-CCAGC CCTTA GCTCA CCTCT G-3 '

EXAMPLE 8

Example 8 sets forth PCR (PCR) conditions, primer sequences, band patterns and enzyme reaction conditions for diagnosing mutations in the toxic gene.

DRD2-Taq1A (Addiction Gene)

※ primer sequence

F: 5'-GACGG CTGGC CAAGT TGTCT A-3 '

R: 5'-GTCGA CCCTT CCTGA GTGTC-3 '

※ Taq I 65 ℃ 2 hour 2% agarose gel

※ enzyme reaction

PCR product 12 ul

Enzyme ul

EXAMPLE 9

Example 9 sets forth PCR (PCR) conditions, primer sequences, band patterns and enzyme reaction conditions for diagnosing mutations in the toxic gene.

DRD2-Taq1B (Addiction Gene)

※ primer sequence

F: 5'-GATGA TACCC ACTTC AGGAA G-3 '

R: 5'-GATGT GTAGG AATTA GCCAG G-3 '

※ Taq I 65 ℃ 2 hour 2% agarose gel

※ enzyme reaction

PCR product 12 ul

Enzyme ul

EXAMPLE 10

Example 10 sets forth PCR (PCR) conditions and enzyme reaction conditions for diagnosing mutations in the curiosity gene.

DRD2-Intron 6 (Curiosity Gene)

※ primer sequence

F: 5'-GGACA GGGGC AATCC TGCAG-3 '

R: 5'-GTCGG GGAGA GTCAG CTGGT-3 '

※ HphI 37 ℃ 2 hour 2% agarose gel

※ enzyme reaction

PCR product 12 ul

Enzyme 0.7ul

EXAMPLE 11

Example 11 sets forth PCR (PCR) conditions and enzyme reaction conditions for diagnosing mutations in the curiosity gene.

DRD4 (Curious Gene)

※ primer sequence

F: 5'-GCTGC TCTAC TGGGC CACGT-3 '

R: 5'-GACCC TCATG GCCTT GCGCT-3 '

EXAMPLE 12

Example 12 presents PCR (PCR) conditions and band patterns for diagnosing mutations in the depressive and violent genes.

Serotonin-Transporter (Depression and Violence Genes)

※ primer sequence

F: 5'-GGCGT TGCCG CTCTG AATGC-3 '

R: 5'-GAGGG ACTGA GCTGG ACAAC CAC-3 '

Therefore, the present invention by the above test method is able to grasp various useful information through the genetic reconstitution personality tests such as genetic diseases or toughness for each individual using DNA genetic factors extracted from the test subject It has a wide range of uses and practical values, which can be widely used as a basis for judgment for good social life as well as for prevention and treatment.

On the other hand, the present invention is not limited to the above embodiments, it is very obvious that various modifications can be presented in the art without departing from the scope of the claims.

Claims (15)

In the test, which is a fluid, After selecting any one of the primer sequences produced by the genetic factors for genetic disease or toughness test, and performing the conditions of the polymerase chain reaction (PCR) so that the presence or absence of genetic mutation of each primer, and the corresponding product Genetic test method for identifying individual characteristics of human genetic factors characterized in that the size of the genotype band can be identified by checking the size of the agarose gel electro-interlocking. The method of claim 1, Genetic factors for constructing the primers of the genetic disease include dementia gene (apoE), low back pain gene (HLA-B27), obesity gene (β-ARG), bone polyvalent electron (VDR), alcohol resistance gene (ADLH2), diabetes gene (IRSII), lung cancer-causing genes (CYPIA1), hypertension genes (AGT) and arthritis genes (ER) genes characterized in that the genetic testing method for the individual characterization of the human genetic factors. The method of claim 1, Genetic factors for constructing the toughness test primers are characterized by including the fitness gene (ACE), addiction gene (DRD2), curiosity genes (DRD2, 4, Intron 6) and depression and violence gene (Serotonin Transporter) Genetic test method for individual characterization of human genetic factors. The method of claim 2, Primer base sequence of dementia gene (apoE), F: 5'-TCG GCC GCA GGG CGC TGA TGG AC-3 ' R: 5'-CCC AGG CGC TCG CGG ATG GCG C-3 ' Inspection condition is, Genetic test method for identifying individual characteristics of human genetic factors characterized in that. The method of claim 2, Primer nucleotide sequence of the back pain gene (HLA-B27), F: 5'-CAGTC TGTGC CTTGG CGTTG C-3 ' R: 5'-GCTAC GTGGA CGACA CGCT-3 ' Inspection condition is, Genetic test method for identifying individual characteristics of human genetic factors characterized in that. The method of claim 2, The primer sequence of the obesity gene (β-ARG), F: 5'-CGC CCA ATA CCG CCA ACA C-3 ' R: 5'-CCA CCA GGA GTC CCA TCA CC-3 ' Inspection condition is, Genetic test method for identifying individual characteristics of human genetic factors characterized in that. The method of claim 2, Primer nucleotide sequence of bone polyvalent electron (VDR), F: 5'-TGC TCA TGG CCA TCT GCA TCG T-3 ' R: 5'-CTA ACC AGC GGA AGA GGT CAA G-3 ' Inspection condition is, Genetic test method for identifying individual characteristics of human genetic factors characterized in that. The method of claim 2, The primer base sequence of the diabetic gene (IRSII), F: 5'-CCA TTA GCA CCC AAA GCT AA-3 ' R: 5'-GTA ATG TGC TAT GTA CGG TA-3 ' Inspection condition is, Genetic test method for identifying individual characteristics of human genetic factors characterized in that. The method of claim 2, Primer nucleotide sequence of arthritis gene (ER), F: 5'-CTC CTT ACA TTC CAT CTG CTG AA-3 ' R: 5'-CAC CCT GGC GTC GAT TAT CTG A-3 ' Inspection condition is, Genetic test method for identifying individual characteristics of human genetic factors characterized in that. The method of claim 3, Primer sequence of the fitness gene (ACE), F: 5'-GCCAC TGCTG GAGAC CACTC-3 ' R: 5'-CCAGC CCTTA GCTCA CCTCT G-3 ' Inspection condition is, Genetic test method for identifying individual characteristics of human genetic factors characterized in that. The method of claim 3, Primer nucleotide sequence of the poisoning gene (DRD2), F: 5'-GACGG CTGGC CAAGT TGTCT A-3 ' R: 5'-GTCGA CCCTT CCTGA GTGTC-3 ' Inspection condition is, Genetic test method for identifying individual characteristics of human genetic factors characterized in that. The method of claim 11, The primer sequence under the same test conditions, F: 5'-GATGA TACCC ACTTC AGGAA G-3 ' R: 5'-GATGT GTAGG AATTA GCCAG G-3 ' Genetic test method for identifying individual characteristics of human genetic factors characterized in that. The method of claim 3, Primer sequence of the curious gene (DRD2), F: 5'-GGACA GGGGC AATCC TGCAG-3 ' R: 5'-GTCGG GGAGA GTCAG CTGGT-3 ' Inspection condition is, Genetic test method for identifying individual characteristics of human genetic factors characterized in that. The method of claim 3, Primer sequence of the curious gene (DRD4), F: 5'-GCTGC TCTAC TGGGC CACGT-3 ' R: 5'-GACCC TCATG GCCTT GCGCT-3 ' Inspection condition is, Genetic test method for identifying individual characteristics of human genetic factors characterized in that. The method of claim 3, The primer sequence of the depression and violent gene is F: 5'-GGCGT TGCCG CTCTG AATGC-3 ' R: 5'-GAGGG ACTGA GCTGG ACAAC CAC-3 ' Inspection condition is, Genetic test method for identifying individual characteristics of human genetic factors characterized in that.