CN107267605B - SNP molecular marker related to Chinese Holstein cow reproduction traits and application thereof - Google Patents
SNP molecular marker related to Chinese Holstein cow reproduction traits and application thereof Download PDFInfo
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Abstract
The invention provides an SNP marker related to Chinese Holstein cow reproductive traits and application thereof, belonging to the technical field of molecular breeding. The invention amplifies a specific fragment from bovine PPARG gene, the nucleotide sequence of the specific fragment is shown as a sequence table SEQ ID NO.1, wherein the 118bp position of the sequence table SEQ ID NO.1 has single base mutation of G → T. The first mating days, nonpregnant days and conception indexes of the Chinese Holstein cow with the base group G on the SNP marker locus are obviously lower than those of the Chinese Holstein cow with the base group T. The invention also provides a primer for detecting the SNP marker and a kit containing the primer. The invention also provides the application of the SNP marker in identifying the dominant variety of the Chinese Holstein cow with high reproductive performance and the application of the SNP marker in the molecular marker assisted breeding of the Chinese Holstein cow, and the SNP marker has the advantages of simple operation, high sensitivity, strong accuracy and low detection cost.
Description
Technical Field
The invention relates to the technical field of molecular biology, in particular to an SNP molecular marker related to Chinese Holstein cow reproductive traits, a kit for detecting the molecular marker, and application of the SNP molecular marker and the kit.
Background
The molecular Marker Assisted Selection (MAS) makes full use of genetic Marker, phenotype and pedigree information, has larger information amount than that of a conventional genetic evaluation method, and can improve the accuracy of individual genetic evaluation. Meuwissen et al (2001) demonstrated by simulations that the accuracy of predicting breeding values based on marker information could reach 85%. In the case of asian assay records, it is of great significance that the estimates of breeding values achieve such high accuracy. The application of MAS in cattle breeding has many advantages: is not easily influenced by the environment, has no limitation of gender and age, allows early seed selection, shortens the generation interval, improves the selection strength, and further improves the efficiency and the accuracy of seed selection. Kashi et al (2008) indicated that the trait gene screening of replacement bulls by MAS in dairy cows was 20-30% faster than that of the conventional descendant determination method. By selecting breeding bulls in advance, MAS can gain a doubling of genetic progress.
PCR-SSCP (polymerase chain reaction-single strand conformation polymorphism) belongs to a form of Single Nucleotide Polymorphism (SNP). The PCR-SSCP molecular marker is a gene variation detection technology developed on the basis of PCR and DNA single-strand gel electrophoresis technology. The basic principle is as follows: in the neutral polyacrylamide gel electrophoresis, the mobility of a single DNA strand depends on the conformation formed by the single DNA strand, besides the length of the DNA strand. Under non-denaturing conditions, the single DNA strand may self-fold into a conformation with a certain spatial structure, which is determined by the base sequence of the single DNA strand, and its stability is maintained by intramolecular interactions (mainly hydrogen bonds). The single DNA strands of the same length show polymorphism, i.e., Single Strand Conformation Polymorphism (SSCP), due to the change of single strand conformation caused by the difference of sequence and even single base, thereby causing the difference of electrophoretic mobility. As a new DNA sequence variation detection means, the PCR-SSCP method has unique points compared with the traditional gene variation detection method: 1) it can find the base mutation of unknown position in the target DNA segment; takao proves that 90% of single base mutation in the DNA fragment of less than 300bp can be found by SSCP through experiments; 2) the mutant single-stranded DNA with different mobility is separated by polyacrylamide gel electrophoresis, and can be further purified.
The genetic homeostasis of the population is disturbed by the high-intensity selection of the lactation performance, and the negative correlation between the lactation performance and the functional traits causes the degradation of the reproductive performance of the high-yield cow population. It is reported that 15-20% of the cows are forced to be eliminated each year due to breeding problems, and the direct and indirect losses caused by them seriously affect the economic efficiency of the dairy industry. Common indexes for measuring reproductive performance mainly comprise the age of a primiparity month, the first mating days after delivery, the number of barren days, calving intervals, conception indexes and the like, and the heritability of the characters is low and data records are lacked, so that the characters are difficult to be considered in a breeding plan. The candidate genes of reproductive performance are taken as detection objects, the correlation between the polymorphism of the genes and the production performance is analyzed by a modern molecular biology technology and a quantitative genetics method, the genotype or allele which is favorable for production is found, and the genes or the genes which are closely linked with the genes are taken as markers to be applied to seed selection, so that the accuracy of seed selection can be improved, more importantly, the animal can be selected by detecting the genotype at the early stage when the animal cannot show the characters, and the method is obviously more economical and reliable than the traditional method of selecting only according to the phenotypic value.
Reproductive performance of cows is closely related to energy metabolism of organisms, and Negative Energy Balance (NEB) in perinatal period is an important reason for the deterioration of reproductive performance of high-yield cows. Peroxisome Proliferator-Activated Receptor gamma (PPAR γ) is a ligand-Activated nuclear transcription factor encoded by PPARG gene, and plays a key role in regulating and controlling animal energy homeostasis, glycolipid metabolism, immune response and the like. Research shows that PPAR gamma is a key transcription factor connecting energy metabolism and reproductive physiological processes of mammals, and high expression of PPAR gamma in corpus luteum and placenta plays an important role in progesterone generation, embryo implantation and pregnancy maintenance. If the nucleotide sequence of the PPARG gene is changed, the PPARG gene may possibly have influence on the combination of the PPARgamma and a ligand and the activation of covalent modification, reduce the affinity of the PPARgamma/RXR alpha and PPRE upstream of a promoter of a gene related to energy homeostasis, further influence the transcriptional activity and expression of the PPARgamma, disturb the energy homeostasis and insulin sensitivity of the organism, and have influence on the reproductive physiological processes such as oestrus, ovulation, embryonic development and the like.
Disclosure of Invention
The invention aims to provide an SNP molecular marker related to Chinese Holstein cow reproductive traits and application thereof.
Another object of the present invention is to provide a kit for detecting the molecular marker.
It is a further object of the present invention to provide a method for identifying chinese holstein cows with high reproductive performance.
The invention provides an SNP molecular marker related to Chinese Holstein cow reproduction traits, the nucleotide sequence of the SNP molecular marker is shown as SEQ ID NO.1, and the base at the 118bp position of the sequence is G or T.
The SNP molecular marker related to the Chinese Holstein cow reproductive traits is obtained by amplifying the primers shown in SEQ ID NO. 2-3.
The invention provides application of the SNP molecular marker in identifying Chinese Holstein cows with high reproductive performance.
Further, when the locus of the sequence shown by the PPARG gene SEQ ID NO.1 shows the GG genotype, the Chinese Holstein cow to be detected belongs to the Chinese Holstein cow with high reproductive performance.
The invention provides an application of the SNP molecular marker in Chinese Holstein cow molecular marker assisted breeding, which comprises the following steps:
(1) screening the SNP markers related to the Chinese Holstein cow reproductive traits by adopting PCR and sequencing technology;
(2) detecting the genotype of the Chinese Holstein cow to be detected by non-denaturing polyacrylamide gel electrophoresis (PAGE);
(3) and (3) breeding the dominant variety of the Chinese Holstein cow with high reproductive performance according to the genotype, wherein when the molecular marker genotype is GG, the Chinese Holstein cow to be tested is the Chinese Holstein cow with high reproductive performance.
As is understood by those skilled in the art, 3 PCR amplification products of individuals with different PAGE band types can be selected for sequencing after non-denaturing polyacrylamide gel electrophoresis, and the genotypes corresponding to the different PAGE band types can be determined. Other individuals which are not sequenced can directly judge the corresponding genotype according to the PAGE banding pattern, and all individuals do not need to be sequenced.
The invention provides a primer pair for detecting the SNP molecular marker, which is respectively a forward primer F: 5'-TGTAACTCAACCTCCTGTT-3' (SEQ ID NO.2) and reverse primer R: 5'-AAGATGCTGTCAGTGAACT-3' (SEQ ID NO. 3).
The invention provides application of the primer pair shown in SEQ ID NO.2-3 in identification of Chinese Holstein cows with high reproductive performance and application in molecular marker-assisted breeding of the Chinese Holstein cows.
The invention provides application of a kit containing the primer pair in identifying Chinese Holstein cows with high reproductive performance and application in molecular marker-assisted breeding of the Chinese Holstein cows.
The invention also provides a method for identifying the Chinese Holstein cow with high reproductive performance,
(1) extracting the genome DNA of the Chinese Holstein cow to be detected;
(2) amplifying a 323bp fragment of the PPARG gene of the Chinese Holstein cow by using the genome DNA of the step (1) as a template and utilizing the primer pair of claim 6 through PCR reaction;
(3) and detecting a PCR amplification product, wherein if the base at the 118bp position in the amplification sequence is G and the genotype is GG, the cattle to be detected belongs to the Chinese Holstein cow with high reproductive performance.
In step (2) of the above method, the amplification system used in the PCR reaction was 25. mu.L: 1 μ L of template DNA, 2.5 μ L of 10 XBuffer (15mmol/L MgCl2), 2 μ L of dNTP (2.5mmol/L), 0.5 μ L of each of primers F and R (12.5pmol/μ L), 0.5 μ L of Taq DNA polymerase (2.5U/μ L), and 18.0 μ L ultrapure water; the PCR reaction conditions in the step (2) are as follows: pre-denaturation at 95 ℃ for 3 min; denaturation at 94 ℃ for 30s, annealing at 58 ℃ for 30s, extension at 72 ℃ for 45s, and 35 cycles; finally, extension is carried out for 10min at 72 ℃.
The SNP loci of the PPARG gene of Chinese Holstein cows are subjected to genotyping, and are subjected to correlation analysis with the first postnatal mating days, nonpregnant days and conception indexes, and SPSS software general linear model program analysis finds that the first postnatal mating days, nonpregnant days and conception indexes of GG genotype individuals are obviously lower than those of TT genotype individuals (P < 0.05). Replacement of the T allele by each G allele resulted in a reduction of 8.83 days for the first postpartum mating, 15.7 days for the barren days, and a 0.25 reduction in conception index. The detection of the polymorphic site provides scientific basis for marker-assisted selection of Chinese Holstein cow reproductive traits.
The SNP marker related to the Chinese Holstein cow reproductive traits and the application thereof have the following advantages:
(1) the molecular genetic marker provided by the invention is not limited by the age and sex of the Chinese Holstein cow, can be used for the early breeding of the Chinese Holstein cow, can accurately screen the Chinese Holstein cow even at the moment of birth, and can remarkably accelerate the genetic progress of the reproductive traits of the Chinese Holstein cow.
(2) The method for detecting the PPARG gene mononucleotide polymorphism of the Chinese Holstein cow has the advantages of accuracy, reliability, simple and convenient operation, high sensitivity and low detection cost.
(3) The detection of the SNP locus of the PPARG gene of the Chinese Holstein cow provides scientific basis for the marker-assisted selection of the propagation traits of the Chinese Holstein cow.
Drawings
FIG. 1 is a SNP locus PAGE typing chart of PPARG gene of Chinese Holstein cows.
FIG. 2 is a graph of sequencing peaks for three genotypes; GG type, TT type and GT type are respectively arranged in sequence.
Detailed Description
The following examples further illustrate the present invention but are not to be construed as limiting the invention. Modifications or substitutions to methods, procedures, or conditions of the invention may be made without departing from the spirit and scope of the invention.
Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.
Example 1 determination of SNP markers associated with the reproductive traits of Chinese Holstein cows and establishment of a method for detecting the same
1. Extracting genome DNA in blood of Chinese Holstein cow to be detected
Blood samples of 293 Chinese Holstein cows in different pastures are taken from jugular vein, ACD is used for anticoagulation, and the blood is stored at-20 ℃ for later use. The genomic DNA in the bovine blood exists in a centrifuge tube solution and is stored at 4 ℃ for standby or stored at-20 ℃ for a long time.
2. Amplification of nucleotide fragments containing SNP sites
Primers were designed based on the PPARG gene (GenBank No. nc — 007320.6) sequences deposited in the NCBI database, including forward primer F: 5'-TGTAACTCAACCTCCTGTT-3' and reverse primer R: 5'-AAGATGCTGTCAGTGAACT-3', using the genome DNA as a template, amplifying the nucleotide fragment of the SNP to be detected, as shown in SEQ ID NO. 1. The SNP site is positioned at 118bp of the PCR amplified fragment, and the base of the site is G or T.
Wherein, the amplification system used in the PCR reaction is 25 μ L: 1 μ L of template DNA, 2.5 μ L of 10 XBuffer (15mmol/LMgCl2), 2 μ L of dNTP (2.5mmol/L), 0.5 μ L of each of primers F and R (12.5pmol/μ L), 0.5 μ L of Taq DNA polymerase (2.5U/μ L), and 18.0 μ L ultrapure water; the PCR reaction conditions are as follows: pre-denaturation at 95 ℃ for 3 min; denaturation at 94 ℃ for 30s, annealing at 58 ℃ for 30s, extension at 72 ℃ for 45s, and 35 cycles; finally, extension is carried out for 10min at 72 ℃.
3. PAGE typing
The genotypes of all Chinese Holstein cows to be detected are detected by a non-denaturing polyacrylamide gel electrophoresis (PAGE) method. The different PAGE banding patterns represent different genotypes, and the typing results of the three genotypes are shown in FIG. 1.
4. Genotype determination
In this example, 3 PCR products of different PAGE banding patterns were selected and sequenced, and the genotypes represented by the different PAGE banding patterns were determined to be GG, TT or GT, respectively, based on the sequencing results. Other individuals that are not sequenced can be directly genotyped from the PAGE banding pattern without sequencing all individuals. If the base at the 118bp position in the amplification sequence is G and the genotype is GG, the Chinese Holstein cow to be detected belongs to the Chinese Holstein cow dominant variety with high reproductive performance. The sequencing peak patterns of the three genotypes are shown in FIG. 2.
5. Application of SNP marker in breeding of Chinese Holstein cow dominant variety
The SNP can be used as a molecular genetic marker of the high reproductive performance of the Chinese Holstein cow, individuals with the genotype of GG are selected and retained, the genotype selection or marker-assisted selection is directly carried out on the Chinese Holstein cow, and a core group of the Chinese Holstein cow with the high reproductive performance is established, so that the breeding of dominant varieties of the Chinese Holstein cow is accelerated.
Example 2 correlation analysis and detection application of different genotypes and reproductive traits of Chinese Holstein cows
The PCR-SSCP assay was performed on 293 Chinese Holstein cows according to the method of example 1, and the results of the analysis of the 118bp locus of the sequence of the PPARG gene shown in SEQ ID NO.1 are shown in Table 1.
TABLE 1 analysis of genetic polymorphisms of SNP loci in the Chinese Holstein cow population
As can be seen from Table 1, the GG type individuals had the highest frequency in the test population, with G being the dominant allele. He was 0.4147 and PIC 0.3270, in the medium polymorphic state. Chi obtained by Pearson's Chi-square test2Is 19.71>χ0.012(df is 2) 9.21, then P<0.01, the test population deviates significantly from Hardy-Weinberg equilibrium at this site, suggesting that its mutations are affected by drift, selection and introduction.
Using SPSS software, calling a general linear model program, analyzing the correlation between the genotype of the SNP polymorphic site and the propagation character, and adopting a model: y isijkl＝μ+Gi+Bj+Yk+MlWherein Y is a phenotypic value of a certain individual trait, μ is a mean value of a certain population trait, GiIs a gene
Type effect, BjFor variety effect, YkFor dating effects, MlIs the milk yield effect and is the random residual effect. The analysis results are shown in tables 2 and 3.
TABLE 2 correlation of SNP loci of PPARG genes of Chinese Holstein cows with reproductive traits (F test)
As can be seen from table 2, genotype significantly affected the first mating days after delivery (P <0.01), days without pregnancy (P <0.05) and conception index (P < 0.05).
Allelic additive, dominant and surrogate effects
Allele additive effects were calculated using the mean of 2 homozygous deviations:
allelic dominant effects were calculated using the dispersion of the mean of heterozygotes from 2 homozygotes:
mean effect of G allele instead of T allele: s ═ a + (q-p) d
Wherein p is the G allele frequency, q is the T allele frequency, and GG, TT and GT are the least square mean of the phenotypes of the genotypes respectively. The significance test for the allele substitution effect was calculated by regression of the reproductive traits to the T allele copy number (0, 1, 2) using a linear regression model (SPSS), defining genotypes TT, GT, and GG as 2, 1, and 0, respectively.
TABLE 3 least square mean value and standard error of PPARG gene of different genotypes of Chinese Holstein cow
In table 3, the same row of the same number has obvious difference in the form of different capital letters, and different capital letters have obvious difference; indicates significant difference (P <0.05) and indicates very significant difference (P < 0.01).
As can be seen from Table 3, the number of days of first mating after GG type delivery is significantly lower than that of GT type (P <0.05) and TT type (P < 0.01); the number of days of nonpregnant GG type is significantly lower than that of GT type (P < 0.05); the GG type 1 subtropical index is significantly lower than the GT type (P <0.01), and the 2 nd subtropical index is significantly lower than the GT type and the TT type (P < 0.05).
For the G allele, the additive effects of the first postnatal mating days, days of nonpregnant and conception index were-6.97 d, -11.2d and-0.215 t, respectively. The gene replacement effect is-8.83 d, -15.70d and-0.25T respectively, namely, the replacement of the T allele by each G allele can lead to 8.83 days of first mating days after delivery, 15.7 days of non-pregnant days and 0.25 time reduction of conception index. Allele G is a high reproductive performance dominant gene.
Example 3 Assembly and application of kit for identifying Chinese Holstein cows with high reproductive performance
The kit comprises 100ng/μ L of bovine blood DNA, upstream and downstream primers with sequences shown in SEQ ID NO.2-3, 2.5mmol/L dNTPs, 2.5U/μ L Taq DNA polymerase, and 10 × Taq Buffer (containing 15 mmol/LMgCl)2) (ii) a Ultrapure water.
F：5′-TGTAACTCAACCTCCTGTT-3′(SEQ ID NO.2)
R：5′-AAGATGCTGTCAGTGAACT-3′(SEQ ID NO.3)
The total PCR reaction system in the kit was 25. mu.L, in which the genomic DNA was 1. mu.L, 10 × buffer (15 mmol/LMgCl)2) 2.5. mu.L, 2. mu.L of dNTP (2.5mmol/L), 0.5. mu.L of each of upstream and downstream primers (12.5 pmol/. mu.L), 0.5. mu.L of Taq DNA polymerase (2.5U/. mu.L), ddH2O18.0. mu.L. PCR reaction procedure: pre-denaturation at 95 ℃ for 3 min; denaturation at 94 ℃ for 30s, annealing at 58 ℃ for 30s, extension at 72 ℃ for 45s, and 35 cycles; finally, extension is carried out for 10min at 72 ℃.
The product obtained by the reaction is the target nucleotide fragment, the sequence is shown as SEQ ID NO.1, the base on the 118bp site of the sequence is detected, and the genotype of the sample is judged. The three genotypes can be classified into GG type, TT type and GT type, if the base at the 118bp site in the amplification product sequence is G, and the genotype is GG type, the Chinese Holstein cow to be detected belongs to the Chinese Holstein cow dominant variety with high reproductive performance. The primers are adopted to carry out PCR amplification on DNA to be detected, amplification products are subjected to non-denaturing polyacrylamide gel electrophoresis (PAGE), PCR amplification products with different PAGE band types are selected for sequencing, and genotypes corresponding to the different PAGE band types are determined. Other individuals which are not sequenced can be directly compared according to the PAGE banding patterns of the determined genotypes, all individuals do not need to be sequenced, and the genotype of GG type, TT type and GT type corresponding to different PAGE banding patterns can be judged, so that the reproductive performance of the individual to be detected can be judged, and the reproductive performance is GG type > GT type > TT type from high to low.
Although the present invention and the embodiments thereof have been described in detail, it should be understood that modifications to the corresponding conditions and the like can be made by those skilled in the art without departing from the technical principle of the present invention, and the modifications should be construed as the scope of the present invention.
SEQUENCE LISTING
<110> national institute of teachers and schools of Gansu province
<120> SNP molecular marker related to Chinese Holstein cow reproduction traits and application thereof
<130>KHP171113296.1
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<170>PatentIn version 3.5
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cctcagacag attgtgacag aacacgtgca gctgttgcaa gtaataaaga aaacagagac 240
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Claims (3)
1. An application of SNP molecular marker in identifying Chinese Holstein cow with high reproductive performance, the nucleotide sequence of the molecular marker is shown as SEQ ID NO.1, the base at the 118bp position of the sequence is G or T;
when the base at the 118bp position of the sequence shown in SEQ ID NO.1 is G and the genotype is GG, the Chinese Holstein cow to be detected belongs to the Chinese Holstein cow with high reproductive performance.
2. The application of a primer pair for detecting a specific SNP molecular marker or a kit containing the primer pair for detecting the specific SNP molecular marker in identifying the Chinese Holstein cow with high reproductive performance is disclosed, wherein the nucleotide sequence of the primer pair is shown as SEQ ID NO.2-3, the nucleotide sequence of the SNP molecular marker is shown as SEQ ID NO.1, and the base at the 118bp position of the SEQ ID NO.1 sequence is G or T.
3. A method for identifying Chinese Holstein cows with high reproductive performance is characterized in that,
(1) extracting the genome DNA of the Chinese Holstein cow to be detected;
(2) taking the genome DNA in the step (1) as a template, and amplifying a PPARG gene 323bp fragment of the Chinese Holstein cow by utilizing a primer pair through PCR reaction; the nucleotide sequence of the primer pair is shown as SEQ ID NO. 2-3;
(3) and detecting a PCR amplification product, wherein if the base at the 118bp position in the amplification sequence is G and the genotype is GG, the cattle to be detected belongs to the Chinese Holstein cow with high reproductive performance.
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| CN112280872A (en) * | 2020-11-18 | 2021-01-29 | 福建农林大学 | SNP molecular marker related to magnesium ion concentration of serum of Chinese Holstein cow |
| CN112210613A (en) * | 2020-11-18 | 2021-01-12 | 福建农林大学 | SNP molecular marker related to serum insulin concentration of Chinese Holstein cow |
| CN113416789B (en) * | 2021-07-27 | 2022-06-03 | 山东省农业科学院畜牧兽医研究所 | Molecular marker related to concentration of beta-hydroxybutyric acid in milk cow blood and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100818052B1 (en) * | 2006-11-21 | 2008-03-31 | 상지대학교산학협력단 | Development of molecular dna marker associated with meat quantity traits of hanwoo |
| CN102168136A (en) * | 2011-02-11 | 2011-08-31 | 中国农业大学 | Application of LHCGR gene of Chinese Holstein cow used as molecular marker |
-
2017
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100818052B1 (en) * | 2006-11-21 | 2008-03-31 | 상지대학교산학협력단 | Development of molecular dna marker associated with meat quantity traits of hanwoo |
| CN102168136A (en) * | 2011-02-11 | 2011-08-31 | 中国农业大学 | Application of LHCGR gene of Chinese Holstein cow used as molecular marker |
Non-Patent Citations (3)
| Title |
|---|
| A missense mutant of the PPAR-γ gene associated with carcass and meat quality traits in Chinese cattle breeds;Yue Yuan Fan;《Genetics and Molecular Research》;20120817;第11卷(第4期);第3781页摘要，第3783页第3段，第3784-3486页，第3787页第2段，图1-3，表1 * |
| ss1006551657;ss1006551657;《dbSNP》;20140523;第1页 * |
| Three novel SNPs in the coding region of PPARc gene and their associations with meat quality traits in cattle;Yue Yuan Fan等;《Molecular biology reports》;20111231;第38卷;第131-137页 * |
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| CN107267605A (en) | 2017-10-20 |
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