KR101763949B1 - Genetic polymorphic markers for determining type of moisture skin and use thereof - Google Patents

Abstract

The present invention relates to a skin type moisturizing diagnostic marker comprising one or more single base polymorphic markers selected from single nucleotide polymorphism (SNP) markers, a composition comprising an agent capable of detecting said marker, a kit comprising said composition Or a microarray and a method for providing information on whether or not the skin type is moisturized using the marker. The single nucleotide polymorphic markers of the present invention provide accurate information on the individual's skin type and thus can develop personalized cosmetics and develop customized active ingredients.

Description

[0002] Moisturizing skin type genetic polymorphism markers and their uses {Genetic polymorphic markers for determining type of moisture skin and use thereof}

The present invention relates to a skin type moisturizing diagnostic marker comprising one or more single base polymorphic markers selected from single base polymorphism (SNP) markers capable of judging whether or not moisturizing skin is present, and a preparation capable of detecting the marker A kit, or a microarray containing the composition, and a method for providing information on whether or not the skin type is moisturized using the marker.

Human skin has different skin condition for each individual. In particular, even skin types of the same age and the same skin type can vary greatly depending on the internal and external environment of each individual. As the scientific civilization develops, there are more and more cases in which the skin is severely affected by exposure to various polluted environments, exposure to ultraviolet rays due to destruction of the ozone layer, various stresses, and the like, resulting in deterioration of skin function and even skin troubles . Accordingly, there is a growing demand for functional cosmetics that can improve or maintain the condition of the skin.

However, at present, it is difficult to obtain complete trust in the skin condition information because the judgment of the skin condition for each user is mainly about the skin condition of the user through simple skin test and simple skin test. In addition, functional cosmetics currently on the market are not aimed at individuals, but are universal. Therefore, there are many limitations in finding suitable cosmetics for users. Therefore, there is almost no system to provide customized cosmetics by precisely diagnosing skin condition by approaching scientific basis.

With the increasing demand for customized cosmetics according to skin characteristics, it is required to establish a systematic standard of skin classification and to characterize product prescription accordingly. However, the current water balance classification standard used for skin classification is not able to accurately classify the skin. In addition, in a method of providing customized cosmetics by measuring the skin condition of the prior art, in order to check the current skin condition of the user, the condition of the skin wrinkles is grasped by an image analyzer system using mainly a replica plate in case of wrinkles, In the case of skin whitening, a change in erythema or skin color is measured using a mexameter and a colorimeter. In the case of skin moisturizing, a moisturizing amount is measured mainly using a corneometer have. These measurement methods simply represent the physical state of the skin surface by numerical values and may vary depending on the program to be measured. Therefore, there is a fear that errors may occur in evaluating the degree of improvement of the subject. In addition, in the case of the visual evaluation of the tester and the evaluation of the questionnaire by the tester, the subjective aspect greatly affects and it is difficult to evaluate the subject's skin condition objectively and precisely.

Under these circumstances, the present inventors constructed a scientific skin classification standard by grasping the genetic characteristics that determine the skin characteristics of an individual, developed a personalized custom active ingredient on the basis thereof, developed customized cosmetics according to skin characteristics through various product segmentation (SNP) markers having a significant correlation with skin moisturization were selected to confirm whether or not the skin type was moisturized. As a result, the present inventors completed the present invention.

It is an object of the present invention to provide a marker for diagnosing whether or not a skin type is moisturized, which comprises at least one single base polymorphism marker selected from single base polymorphism (SNP) markers capable of judging whether or not the skin is moisturized.

Another object of the present invention is to provide a composition for diagnosing whether or not a skin type is moisturized, which comprises a probe capable of detecting a skin-type moisturizing or non-moisturizing marker or an amplifiable preparation.

Another object of the present invention is to provide a skin type moisturizing kit or microarray comprising the skin type moisturizing composition.

Yet another object of the present invention is to provide a method for providing information on whether or not a skin type is moisturized, which comprises identifying a polymorphic site of the single nucleotide polymorphism marker.

In accordance with one aspect of the present invention, the present invention provides a skin type skin moisturizing diagnostic marker comprising at least one single base polymorphism marker selected from single base polymorphism (SNP) to provide.

In the present invention, the term "polymorphism" refers to a case where two or more alleles exist in one locus. Of the polymorphic sites, only a single base differs from a polymorphism region to a single base polymorphism (single nucleotide polymorphism, SNP). Preferred polymorphic markers have two or more alleles exhibiting an incidence of 1% or more, more preferably 10% or 20% or more, in the selected population.

The term "allele " in the present invention refers to various types of genes that exist on the same locus of a homologous chromosome. Alleles are also used to represent polymorphisms, for example, SNPs have two kinds of bialles.

In the present invention, the term "rs_id " means rs-ID, which is an independent marker assigned to all SNPs initially registered by the NCBI that has started accumulating SNP information since 1998. [ The rs_id shown in the above table means the SNP marker which is the polymorphism marker of the present invention.

In the present invention, the term "skin type " refers to a skin type of an individual to be measured or diagnosed. The type of skin that can be measured by the SNP of the present invention is not limited but may be moisturized. According to one embodiment of the present invention, the inventors classified the subject's skin type into a moist skin (moist skin) or a moist skin. Since the single nucleotide polymorphic marker of the present invention enables accurate skin type measurement, information on the changed skin type of skin contacted with the active ingredient can also be provided.

Preferably, the single nucleotide polymorphism marker may be at least one single nucleotide polymorphism marker selected from the single nucleotide polymorphism markers set forth in Table 3 and Table 4. The single nucleotide polymorphism markers shown in Table 3 and Table 4 may be used to determine whether the skin type is moisturizing.

Whether the single nucleotide polymorphism marker of the present invention was diagnosed with the skin type was determined by measuring the frequency of each marker. Such significance is not limited to p-values such as less than 0.05, less than 0.01, less than 0.001, less than 0.0001, less than 0.00001, less than 0.000001, less than 0.0000001, less than 0.00000001, or less than 0.000000001 p-value. Preferably, the p-value may be less than 0.01, more preferably the p-value may be less than 0.001.

The polymorphic markers of the present invention are the markers shown in Table 3 and Table 4 when the polymorphic site of the individual polymorphic marker is a minor allele. ) And the frequent frequency of the watery skin test case is judged to be the skin of the high frequency group. If the polymorphic base is a major allele, the water shown in Table 3 and Table 4 is low This is a marker that can be judged as the skin of the lower frequency group in the frequency of the test group which is the skin control group and the watery skin group. For example, when the 129412726 base of the chromosome 3 of the individual is T, the frequency of the control group is higher than that of the control group. Therefore, the skin of the individual having the minority allele T is judged as a control skin You can. If the 129412726 base of chromosome 3 of the individual is C, which is the majority allele of the individual, the frequency of the test group is lower than that of the test group.

More preferably, Table 3 shows a polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 129412726 base of human chromosome 3, C or T (rs28459381), the 129412726 base; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 29663843th base of human chromosome 6, C or T (rs1233387), the 29663843th base; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 29820388 base of the human chromosome 7, C or T (rs174922), and the 29820388 base; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 29820850 base of the human chromosome 7, T or C (rs174923), and the 29820850 base; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising 141425903 base of human 7 or C14 or base 15142903 base (rs12666982); A polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 29105682 base of human chromosome 3 is G or A (rs9818278), and the 29105682 base; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 125175173 base of the human chromosome 5, T or C (rs12109492), including the 125175173 base; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising 59132422 bases of human chromosome 12 (rs12228176), which is C or T, and 59132422 bases; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 52868639 base of the human chromosome 2, C or T (rs7581434), and the 52868639 base; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 5285619595 base of human human chromosome 2, G or T (rs7573204), and the 52856195 base; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 59106853 base of human chromosome 12, T or C (rs3923917), and the 59106853 base; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 4600440 base of the human chromosome 20 (A rs28702443) and the 4600440 base; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 29914639 base of the human tenth chromosome is G or A (rs4572039), and the 29914639 base; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 95705228 base of the human eighth chromosome (rs2890805), the 95705228 base being C or T; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 25590529 base of the human 15th chromosome (rs25590529), which is A or G, and the 25590529 base; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 107093501 base of the human 6 th chromosome, G or A (rs783400), the 107093501 base; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 25586993 base of the human 15th chromosome (rs3098582) with C or T, the 25586993 base; A polynucleotide consisting of 5-100 consecutive DNA sequences comprising the 25585078 base of the human 15th chromosome (rs12591822) with T or C; And at least one polynucleotide selected from the group consisting of complementary polynucleotides thereof. The skin type may be a moisturizing or non-moisturizing skin marker for determining whether the skin type is moist or too moist.

The present inventors confirmed that the above-mentioned SNP is a skin type diagnostic marker through the following proof. Specifically, the moisturizing skin condition is a skin control with a CM value of 30 or less based on the water content measured using a water meter, Corneometer CM825 (C + K, Germany), a moist skin test with a CM value of 60 or more Respectively. GDNA was extracted from the saliva or blood from the classified individuals and genotype analysis experiments were performed with an Axiom (TM) ASI array plate (Fig. 1). As a result, SNPs having significance in moisturizing skin type were classified (Table 3 and Table 4). SNPs that can classify whether such skin types are moisturized are those first identified by the present inventors.

In another aspect, the present invention provides a composition for diagnosing whether or not a skin type moisturizes, comprising a probe capable of detecting the skin type moisturizing diagnostic marker or an agent capable of amplifying the skin type.

In the present invention, the term "a probe capable of detecting a skin type moisturizing diagnostic marker" refers to a composition capable of diagnosing whether a skin type is moisturized by specifically hybridizing with a polymorphic site of the above- And the specific method of such gene analysis is not particularly limited and may be by any gene detection method known in the art to which this invention belongs.

In the present invention, the term "agent capable of amplifying a skin type moisturizing diagnostic marker" means a composition capable of diagnosing whether a skin type is moisturized by confirming a polymorphic site of such a gene through amplification, Preferably means a primer capable of specifically amplifying the polynucleotide of the marker for diagnosing whether or not the skin type is moisturized.

The primers used for the polymorphic marker amplification can be amplified by PCR using appropriate conditions in suitable buffers (e.g., four different nucleoside triphosphates and polymerase such as DNA, RNA polymerase or reverse transcriptase) and template-directed DNA Refers to a single stranded oligonucleotide that can serve as a starting point for synthesis. The appropriate length of the primer may vary depending on the intended use, but is usually 15 to 30 nucleotides. Short primer molecules generally require a lower temperature to form a stable hybrid with the template. The primer sequence need not be completely complementary to the template, but should be sufficiently complementary to hybridize with the template.

The term "primer" in the present invention means a base sequence having a short free 3 'hydroxyl group and can form base pairs with a complementary template, It means a short sequence functioning as a point. The primer can initiate DNA synthesis in the presence of reagents and four different nucleoside triphosphates for polymerization reactions (i.e., DNA polymerase or reverse transcriptase) at appropriate buffer solutions and temperatures. PCR amplification can be performed to predict skin type through the production of desired products. The PCR conditions, the lengths of the sense and antisense primers can be modified based on what is known in the art.

The probes or primers of the present invention can be chemically synthesized using the phosphoramidite solid support method, or other well-known methods. Such nucleic acid sequences may also be modified using many means known in the art. Non-limiting examples of such modifications include, but are not limited to, methylation, "capping ", replacement of natural nucleotides with one or more homologues, and modifications between nucleotides, such as uncharged linkers, such as methylphosphonate, Phosphoamidates, carbamates, etc.) or charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.).

In another aspect, the present invention provides a skin type moisturizing diagnostic kit comprising the composition for diagnosing whether or not the skin type is moisturized.

The kit may be an RT-PCR kit or a DNA chip kit.

The kit of the present invention can diagnose the skin type by confirming the SNP polymorphism marker which is a skin type diagnostic marker through amplification or by confirming the expression level of SNP polymorphism marker with the mRNA expression level. As a specific example, in the present invention, the kit for measuring the mRNA expression level of the skin type diagnostic marker may be a kit containing essential elements necessary for performing RT-PCR. In addition to the individual primer pairs specific for the genes of the skin type diagnostic marker, the RT-PCR kit also includes a test tube or other appropriate container, reaction buffer (pH and magnesium concentrations vary), deoxynucleotides (dNTPs) , Enzymes such as Taq polymerase and reverse transcriptase, DNase, RNAse inhibitors, DEPC-water, sterile water, and the like. It may also contain a primer pair specific for the gene used as a quantitative control. Also preferably, the kit of the present invention may be a skin type diagnostic kit containing essential elements necessary for carrying out a DNA chip. DNA chip kits are those in which nucleic acid species are attached in a gridded array on a generally flat solid support plate, typically a glass surface not larger than a slide for a microscope, and nucleic acids are uniformly arranged on the chip surface, Hybridization reaction occurs between the nucleic acid on the surface and the complementary nucleic acid contained in the solution treated on the surface of the chip to enable a mass parallel analysis.

In another aspect, the present invention provides a microarray for diagnosing whether or not a skin type moisturizing condition includes a polynucleotide of a skin-type moisturizing diagnostic marker.

The microarray may comprise DNA or RNA polynucleotides. The microarray comprises a conventional microarray except that the polynucleotide of the present invention is contained in the probe polynucleotide.

Methods for producing microarrays by immobilizing probe polynucleotides on a substrate are well known in the art. The probe polynucleotide means a polynucleotide capable of hybridizing, and means an oligonucleotide capable of binding to the complementary strand of the nucleic acid in a sequence-specific manner. The probe of the present invention is an allele-specific probe in which a polymorphic site exists in a nucleic acid fragment derived from two members of the same species and hybridizes to a DNA fragment derived from one member but does not hybridize to a fragment derived from another member . In this case, the hybridization conditions show a significant difference in the intensity of hybridization between alleles, and should be sufficiently stringent to hybridize to only one of the alleles. This can lead to good hybridization differences between different allelic forms. The probe of the present invention can detect an allele and can be used for diagnosis of skin type and the like. The diagnostic methods include detection methods based on hybridization of nucleic acids such as Southern blots, and may be provided in a form pre-bonded to a substrate of a DNA chip in a method using a DNA chip. The hybridization can usually be performed under stringent conditions, for example, a salt concentration of 1 M or less and a temperature of 25 ° C or higher. For example, conditions of 5x SSPE (750 mM NaCl, 50 mM Na Phosphate, 5 mM EDTA, pH 7.4) and 2530 [deg.] C may be suitable for allele-specific probe hybridization.

Immobilization on the substrate of the probe polynucleotide associated with the skin diagnosis of the present invention can also be easily made using this conventional technique. In addition, hybridization of nucleic acids on a microarray and detection of hybridization results are well known in the art. The detection can be accomplished, for example, by labeling the nucleic acid sample with a labeling substance capable of generating a detectable signal comprising a fluorescent material, such as Cy3 and Cy5, and then hybridizing on the microarray and generating The hybridization result can be detected.

In another aspect, the present invention provides a method for producing a DNA comprising: (a) obtaining DNA from a sample isolated from an individual; (b) amplifying the polymorphic site of the single nucleotide polymorphic marker from the DNA obtained in step (a) or hybridizing with the probe; And (c) identifying the base of the amplified or hybridized polymorphic site of step (b).

The term "individual" of the present invention means a subject for diagnosing whether or not the skin type is moisturizing. DNA can be obtained from the sample, such as hair, urine, blood, various body fluids, isolated tissues, isolated cells or saliva, but is not limited thereto.

The method of obtaining the genomic DNA of step (a) may be any method known to those skilled in the art.

The amplification of the polymorphic site of the single nucleotide polymorphic marker from the DNA obtained in step (a) of step (b) or hybridization with the probe may be performed by any method known to those skilled in the art. For example, the target nucleic acid can be obtained by PCR amplification and purification thereof. Other ligase chain reaction (LCR) (Wu and Wallace, Genomics 4, 560 (1989), Landegren et al., Science 241, 1077 (1988)), transcription amplification (Kwoh et al., Proc. Natl. Acad. Sequence amplification based on nucleic acids (NASBA) can be used as well as self-sustaining sequence replication (Guatelli et al., Proc. Natl. Acad. Sci. USA 87, 1874 (1990)).

Determination of bases in the polymorphic site of step (c) of the above method can be performed by sequencing analysis, hybridization by microarray, allele specific PCR, dynamic allele-specifichybridization, DASH), PCR extension analysis, SSCP, PCR-RFLP analysis or TaqMan technique, SNPlex platform (Applied Biosystems), mass spectrometry (e.g., Sequenom's MassARRAY system), mini-sequencing method, Bio- But are not limited to, the BioRad system, the CEQ and SNPstream system (Beckman), the Molecular Inversion Probe array technology (e.g., Affymetrix GeneChip), and BeadArray Technologies (e.g. Illumina GoldenGate and Infinium assay). One or more alleles in a polymorphic marker, including microsatellite, SNP or other types of polymorphic markers, can be identified by such methods or by other methods available to those skilled in the art to which this invention pertains. The base of such a polymorphic site can be determined preferably through a SNP chip.

The term "SNP chip" in the present invention means one of DNA microarrays capable of confirming each base of several hundred thousand SNPs at a time.

The TaqMan method comprises the steps of: (1) designing and constructing a primer and a TaqMan probe to amplify a desired DNA fragment; (2) labeling probes of different alleles with FAM dyes and VIC dyes (Applied Biosystems); (3) performing PCR using the DNA as a template and using the primer and the probe; (4) after completion of the PCR reaction, analyzing and confirming the TaqMan assay plate with a nucleic acid analyzer; And (5) determining the genotype of the polynucleotides of step (1) from the analysis results.

In the above, the sequencing analysis can be performed using a conventional method for determining the nucleotide sequence, and can be performed using an automated gene analyzer. The allele-specific PCR means a PCR method in which a DNA fragment in which the SNP is located is amplified with a primer set including a primer designed with the base at the 3 'end at which the SNP is located. The principle of the above method is that, for example, when a specific base is substituted by A to G, an opposite primer capable of amplifying a primer containing the A as a 3 'terminal base and a DNA fragment of an appropriate size is designed, In the case where the base at the SNP position is A, the amplification reaction is normally performed and a band at a desired position is observed. When the base is substituted with G, the primer can be complementarily bound to the template DNA, And the amplification reaction is not performed properly due to the inability of complementary binding at the terminal. DASH can be performed by a conventional method, preferably by a method such as Prince et al.

On the other hand, in the PCR extension analysis, first, a DNA fragment containing a base in which a single base polymorphism is located is amplified by a pair of primers, and all nucleotides added to the reaction are deactivated by dephosphorylation, and SNP- specific extension primers, a dNTP mixture, a digoxin nucleotide, a reaction buffer, and a DNA polymerase to perform a primer extension reaction. At this time, the extension primer has a base immediately adjacent to the 5 'direction of the base in which the SNP is located at the 3' terminus, and the nucleic acid having the same base as the dodecoxynucleotide is excluded in the dNTP mixture, and the dodecoxynucleotide indicates the SNP Base type. For example, when dGTP, dCTP and TTP mixture and ddATP are added to the reaction in the presence of substitution from A to G, the primer is extended by the DNA polymerase in the base in which the substitution has occurred, The primer extension reaction is terminated by ddATP at the position where the base first appears. If the substitution has not occurred, the extension reaction is terminated at the position, so that it is possible to discriminate the type of the base representing the SNP by comparing the lengths of the extended primers.

As the detection method, when the extension primer or the dioxynucleotide is fluorescently labeled, the SNP is detected by detecting fluorescence using a gene analyzer (for example, Model 3700 of ABI Co., Ltd.) used for general nucleotide sequence determination And when the unlabeled extension primer and the didyxin nucleotide are used, the SNP can be detected by measuring the molecular weight using MALDI-TOF (matrix assisted laser desorption ionization-time of flight) technique.

Preferably, the method of providing information of the present invention comprises: (d) when the base of the amplified or hybridized polymorphic site is a minor allele, the control group, which is a skin with low water shown in Table 3 and Table 4, The frequency of the skin test group is judged to be the skin of the high frequency group. If the base of the amplified or hybridized polymorphic region is a major allele, the control group, which is a skin with low water content shown in Table 3 and Table 4 And a step of judging the skin of the group having a low frequency frequency frequency of the test group, which is a moist skin, to be low.

The single nucleotide polymorphic marker of the present invention is a marker capable of diagnosing skin characteristics for each individual, and can give information on the accurate skin type to an individual, thus providing a customized cosmetic according to the skin characteristics. In addition, by classifying the skin characteristics of cosmetics consumers scientifically, it is possible to develop tailor-made effective ingredients according to individuals and to develop customized cosmetics according to skin characteristics through various product segmentation.

1 is a schematic diagram of gene analysis using the Axiom (TM) ASI array plate of the present invention.
FIG. 2 is a graph showing the log value of the p-value with respect to the frequency of each SNP marker for moisturizing and the chromosomal location of the SNP.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and the scope of the present invention is not construed as being limited by these embodiments.

Example 1: Characterization of Skin and Gene Collection

In order to establish a skin classification standard for each type of gene and to develop a personalized active ingredient, the present inventors conducted a study on the skin moisturizing, which is an important factor of skin physiological factors, in order to classify the skin characteristics of healthy female subjects of 20 to 50 years old. 81 subjects who met the subject selection criteria of the present invention and 20 subjects who completed whitening skin classification in the dermatology department of Gyeongsang National University were selected from the basic data of the subjects possessed by the pros and then the skin characteristics were classified and samples were collected ). The sample means saliva or blood. For example, the saliva was collected twice, and the first saliva was collected after subjects were fasted one hour before collection, and the second saliva was obtained immediately after the morning wake.

The classification was carried out according to the following criteria.

○ Classification of skin moisturizing (moisture): Classification of skin moisturizing was classified by the moisture amount measured by Corneometer CM825 (C + K, Germany) CM value of 30 or less was used as a skin control group, and CM value of 60 or more was used as a moisturizing skin test group.

Classification The control (control) The test case Moisturizing skin (moisture) CM value less than 30 CM value 60 or higher

If you are pregnant or breastfeeding, or if you plan to become pregnant within 6 months, (2) you have used steroid-containing skins for at least one month to treat skin conditions, (3) you have not taken more than six months ⑤ If you have sensitive or irritable skin, ⑤ If you have skin abnormalities such as dots, acne, erythema and capillary dilatation on the test site. ⑥ You can not apply the same or similar cosmetics or medicines ⑦ If you have a chronic wasting disease (asthma, diabetes, hypertension, etc.), ⑨ If you have atopic dermatitis, ⑩ Other cases In cases where it was judged by the examiner that the examination was difficult, the subjects were excluded.

Example  2: Genotype analysis according to skin characteristics

Genotyping of the skin type samples was performed using the Affymetrix Axiom (TM) ASI array plate.

In order to determine whether the skin type was moisturized according to the criteria of Example 1, the skin type consisted of 20 persons in each of 10 test groups and 10 control groups, and gDNA was extracted from the saliva in the forties.

The saliva or blood of women in each group was extracted with QIAmp mini gDNA prep kit (QIAGEN, Korea) and OD260 / 280> 1.7, more than 10ng / ㎕, 1xTAE 1% agarose gel, After confirming the band, the genotype analysis experiment was carried out. Genomic assays using the Axiom ASI plate were performed in accordance with the manual. The reagents contained in the Axiom ASI plate kit were used for amplification of gDNA, DNA fragmentation, quality control, hybridization, staining, and scanning. The image of the chip was scanned using the GeneTitan MC (Affymetrix) system and the CeneChip Command Console Software (AGCC), and the scanned image dat file was automatically converted into a cel file by the AGCC. The cel file for each chip was subjected to data quality control and genotype call using genotyping console 4 program. A general schematic diagram of a method for gene analysis using the Affymetrix Axiom (TM) ASI array plate is shown in FIG.

A total of 20 genotyping chip experiments were carried out, and all the samples passed QC and genotyping results were obtained. QC standards are passed if the DISHQC value of the Axiom Chip is greater than 0,82 and the genotype call rate is more than 97%.

Example 3: Survey of gene polymorphism of moisturizing skin using genotype analysis of skin type and screening of significant SNP

In order to detect genotypes associated with skin type, association analysis using SNP markers of test group / control group was performed. For this, we used the PLINK 1.07 program and set MAF (Minor Allele Frequency)> 0.1 and Hardy-Weinberg Equilibrium (HWE)> 0.001 as QC parameters for each marker.

The test group / control group were compared, and the number of data used in the comparison was as follows (Table 2). The final major SNP markers predicted to be associated with the skin type selected only the Affymetrix SNP annotation associated with the skin type, with a significance level of the chi-square test for each allele of p <0.0001 or less, or p <0.01.

The test case The control (control) Number of SNPs Moisturizing 10 10 18

In addition, we selected a significant level of chi-square test with respect to moisturizing to be p <0.01.

The log-value of the p-value with respect to the frequency of each marker in the association analysis results related to moisturization and the chromosomal location of the SNP are shown in FIG. As a result, it was confirmed that SNP markers having high significance exist sporadically throughout the chromosome. A list of major SNP markers significantly associated with such moisturization is shown in Table 3 below.

The SNP marker list of p <0.01 significantly associated with moisturization is shown in Table 4 below.

Claims (10)

delete delete A composition for diagnosing highly water-impermeable skin, comprising a probe capable of detecting a single nucleotide polymorphism (SNP) marker for diagnosing whether or not the skin is highly water-soluble, or an amplifiable agent,
The single nucleotide polymorphism marker for diagnosing whether or not the skin is highly water-soluble is composed of 5-100 consecutive DNA sequences containing 129412726 base (Cys or Ts) of human chromosome 3 (rs28459381) and 129412726 base Wherein the composition is composed of a polynucleotide or a complementary polynucleotide of SEQ ID NO:
A kit for diagnosing whether or not a skin is highly water-soluble, comprising the composition of claim 3.
5. The kit according to claim 4, wherein the kit is an RT-PCR kit or a DNA chip kit.
A microarray for diagnosing whether or not a skin is highly water-resistant, comprising a single nucleotide polymorphism (SNP) marker for diagnosing whether or not the skin is highly water-
The single nucleotide polymorphism marker for diagnosing whether or not the skin is highly water-soluble is composed of 5-100 consecutive DNA sequences containing 129412726 base (Cys or Ts) of human chromosome 3 (rs28459381) and 129412726 base Or a complementary polynucleotide of the polynucleotide or its complementary polynucleotide.
(a) obtaining DNA from a sample isolated from an individual;
(b) amplifying or hybridizing a polymorphic site of a single nucleotide polymorphism (SNP) marker for diagnosing whether the skin is highly water-soluble, from the DNA obtained in the step (a)
The single nucleotide polymorphism marker for diagnosing whether or not the skin is highly water-soluble is composed of 5-100 consecutive DNA sequences containing 129412726 base (Cys or Ts) of human chromosome 3 (rs28459381) and 129412726 base &Lt; / RTI &gt; or a complementary polynucleotide thereof; And
(c) identifying the base of the amplified or hybridized polymorphic site of step (b).
8. The method of claim 7, wherein (d) if the base of the amplified or hybridized polymorphic site is T, it is judged to have low-moisture skin, and if the base of the amplified or hybridized polymorphism site is C, The method according to any one of claims 1 to 3, further comprising the step of determining that the skin is highly water-impermeable.
8. The method of claim 7, wherein the sample is hair, urine, blood, various body fluids, isolated tissue, isolated cells or saliva.
8. The method of claim 7, wherein the amplification and confirmation of the polymorphic site uses a SNP chip.

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