CN111850139B - A molecular marker located on porcine chromosome 12 related to the formation of porcine single testis and its application - Google Patents
A molecular marker located on porcine chromosome 12 related to the formation of porcine single testis and its application Download PDFInfo
- Publication number
- CN111850139B CN111850139B CN202010788014.XA CN202010788014A CN111850139B CN 111850139 B CN111850139 B CN 111850139B CN 202010788014 A CN202010788014 A CN 202010788014A CN 111850139 B CN111850139 B CN 111850139B
- Authority
- CN
- China
- Prior art keywords
- pig
- molecular marker
- chromosome
- testis
- formation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000003147 molecular marker Substances 0.000 title claims abstract description 54
- 210000000349 chromosome Anatomy 0.000 title claims abstract description 41
- 230000015572 biosynthetic process Effects 0.000 title claims abstract description 28
- 210000001550 testis Anatomy 0.000 title claims abstract description 18
- 241000282887 Suidae Species 0.000 claims abstract description 30
- 230000002068 genetic effect Effects 0.000 claims abstract description 9
- 230000035772 mutation Effects 0.000 claims abstract description 9
- 230000006872 improvement Effects 0.000 claims abstract description 7
- 241000282898 Sus scrofa Species 0.000 claims description 102
- 238000000034 method Methods 0.000 claims description 23
- 230000009395 genetic defect Effects 0.000 claims description 10
- 108700028369 Alleles Proteins 0.000 claims description 8
- 239000002773 nucleotide Substances 0.000 claims description 8
- 125000003729 nucleotide group Chemical group 0.000 claims description 8
- 206010055002 Monorchidism Diseases 0.000 claims description 5
- 210000000918 epididymis Anatomy 0.000 claims description 5
- 201000010063 epididymitis Diseases 0.000 claims description 5
- 238000012216 screening Methods 0.000 claims description 5
- 238000011144 upstream manufacturing Methods 0.000 claims description 5
- 238000001514 detection method Methods 0.000 claims description 4
- 150000007523 nucleic acids Chemical group 0.000 claims description 4
- 238000012214 genetic breeding Methods 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 claims 1
- 238000009395 breeding Methods 0.000 abstract description 17
- 230000001488 breeding effect Effects 0.000 abstract description 17
- 238000005516 engineering process Methods 0.000 abstract description 6
- 238000012098 association analyses Methods 0.000 abstract description 3
- 108090000623 proteins and genes Proteins 0.000 description 9
- 238000004458 analytical method Methods 0.000 description 8
- 230000000694 effects Effects 0.000 description 5
- 238000012163 sequencing technique Methods 0.000 description 5
- 238000012408 PCR amplification Methods 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 239000003550 marker Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000008827 biological function Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000010219 correlation analysis Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 238000012252 genetic analysis Methods 0.000 description 2
- 238000013507 mapping Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 238000003307 slaughter Methods 0.000 description 2
- 238000013517 stratification Methods 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000001712 DNA sequencing Methods 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 208000011622 Testicular disease Diseases 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- YTRQFSDWAXHJCC-UHFFFAOYSA-N chloroform;phenol Chemical compound ClC(Cl)Cl.OC1=CC=CC=C1 YTRQFSDWAXHJCC-UHFFFAOYSA-N 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000012165 high-throughput sequencing Methods 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 235000020997 lean meat Nutrition 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 238000000611 regression analysis Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 210000000689 upper leg Anatomy 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/124—Animal traits, i.e. production traits, including athletic performance or the like
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
本发明属于分子生物技术和分子标记技术领域，具体涉及一种位于猪12号染色体上与猪单睾形成相关的分子标记及应用。所述的位于猪12号染色体上与猪单睾形成相关的分子标记的SNP位点对应于国际猪基因组11.1版本参考序列12号染色体上第26276230位T>C突变。该分子标记是通过全基因组关联分析得到，该分子标记与杜洛克猪单睾性状显著相关。本发明还提供了一种用于鉴定该分子标记的引物对，利用该分子标记和引物对可建立高效准确的分子标记辅助育种技术，将其应用于种猪单睾性状遗传改良中，从而保障企业利润，增加核心竞争力。
The invention belongs to the technical field of molecular biotechnology and molecular markers, in particular to a molecular marker located on pig chromosome 12 and related to the formation of porcine single testis and its application. Said SNP site on the porcine chromosome 12 of the molecular marker related to the formation of porcine monotestis corresponds to the T>C mutation at position 26276230 on chromosome 12 of the reference sequence of the international porcine genome version 11.1. The molecular marker was obtained by genome-wide association analysis, and the molecular marker was significantly associated with the single testis trait of Duroc pigs. The present invention also provides a primer pair for identifying the molecular marker. By using the molecular marker and the primer pair, an efficient and accurate molecular marker-assisted breeding technology can be established and applied to the genetic improvement of the single testis trait of breeding pigs, so as to ensure the protection of enterprises profit and increase core competitiveness.
Description
Technical Field
The invention belongs to the technical field of molecular biotechnology and molecular marker, and particularly relates to a molecular marker located on a pig No. 12 chromosome and related to formation of a pig mono-testis and application thereof.
Background
The slaughtering amount of domestic pigs accounts for more than half of the total slaughtering amount of the domestic pigs in the world, and the output value created by the pig raising industry and the economic benefit generated by the production value have very important influence on the development of the economic society. However, the various genetic defects and diseases of swine often result in significant breeding and economic losses to the swine industry. The pig monorchid character, namely that the piglet only has one testis after birth, and the individual with the genetic defect is eliminated in the breeding process. On one hand, the main reason for causing the character of the pig mono-testis is the genetic factor, and the pig mono-testis is taken as a complex character and is controlled by polygene; on the other hand, for the breeding pig farm, the breeding boar with the genetic defect of the single testis loses the breeding value and is eliminated, which directly causes the economic loss of the breeding pig farm. Therefore, it is very important to identify Quantitative Trait Loci (QTL) affecting the pig monorchid trait. At present, the QTL detection research method for complex economic traits of livestock and poultry comprises three methods: candidate Gene Analysis (CGA), QTL linkage analysis (QTL linkage analysis, also known as QTL mapping), and Genome-wide association analysis (GWAS).
The candidate gene method is to explore the relation between the candidate gene and the target character on the premise that the physiological function and the strong correlation in the growth and development process exist between the candidate gene and the target character. Although the method has the advantages of simple operation and low total cost, the method only aims at the genes with known biological functions and cannot find the QTL with unknown biological functions. With the development of molecular biology, the emergence of QTL mapping based on pedigree and molecular markers has broken this limitation. Compared with a candidate gene method, the QTL positioning method researches the relationship between genetic markers and characters in the whole genome range, and the corresponding position of the QTL in the genome is judged by acquiring the linkage disequilibrium relationship between the genetic markers and the QTL. The QTL positioning method is a relatively precise positioning method, but still has the defects of poor resolution, incapability of accurately positioning the QTL related to the characters at one time, incapability of analyzing a plurality of characters at one time, long period, low efficiency and the like. GWAS is considered as a new approach for high resolution genetic analysis. With the rapid development of genomics, high-throughput sequencing technology, high-density SNP chip technology and computer technology, GWAS gradually replaces QTL positioning method to become an important means for candidate gene identification and genetic analysis of complex traits, and is widely applied to human beings, plants and animals.
The Duroc pig as lean type pig breed has the characteristics of strong adaptability, wide distribution, fast growth speed, high feed conversion efficiency, high carcass lean meat percentage and the like, and is widely used as a terminal male parent of a Duroc (Duroc X long white pig X big white pig) commercial pig. Duroc pigs serve as terminal male parents, all performance indexes of the Duroc pigs have extremely high standards, breeding of the Duroc boars is very important, and the Duroc boars with genetic defects can directly influence the income of a breeding farm. Therefore, the research on the potential pathogenic site causing the single testis character of the core group of the Duroc pigs and the genetic improvement can reduce the great economic loss brought by the genetic defect to the maximum extent to the fastest extent for breeding enterprises and ensure the profit of the enterprises.
Disclosure of Invention
In order to overcome the defects and shortcomings of the prior art, the invention mainly aims to provide a molecular marker which is located on the pig chromosome 12 and is related to the formation of the pig mono-testis.
The invention also aims to provide application of the molecular marker which is positioned on the pig chromosome 12 and is related to the formation of the pig mono-testis.
It is still another object of the present invention to provide a primer set for identifying the above molecular marker.
The fourth object of the present invention is to provide the use of the above primer set.
The fifth purpose of the invention is to provide a genetic improvement method of Duroc pigs.
The purpose of the invention is realized by the following technical scheme:
a molecular marker which is positioned on a chromosome 12 of a pig and is related to the formation of a pig monocrchidism, and the SNP locus of the molecular marker corresponds to the 26276230T > C mutation on the chromosome 12 of a reference sequence version 11.1 of an international pig genome;
the nucleotide sequence of the molecular marker which is positioned on the pig chromosome 12 and is related to the formation of the pig mono-testis is preferably shown as SEQ ID NO:1, wherein M in the sequence is T or C, resulting in a difference in the number of pig testes;
the molecular marker which is positioned on the pig No. 12 chromosome and is related to the formation of the pig mono-testis is applied to the identification of the character and the genetic breeding of the Duroc pig mono-testis;
a method for detecting the characteristics of the pig monorchidism comprises the following steps:
detecting the molecular marker which is positioned on the pig No. 12 chromosome and is related to the formation of the pig single testis on the pig No. 12 chromosome, wherein the single nucleotide of the SNP locus of the molecular marker is T or C;
the pig is preferably a pure Duroc and a synthetic line thereof;
a primer pair for identifying the molecular marker which is positioned on the pig chromosome 12 and is related to the formation of the pig mono-testis comprises a primer-F and a primer-R, and the nucleic acid sequences are as follows:
upstream primer-F: 5'-AGAAGCTGGACTTAGAACCC-3';
downstream primer-R: 5'-GAGCCTGGAAAATCTATCCCT-3';
a kit for identifying the molecular marker which is positioned on the pig chromosome 12 and is related to the formation of the pig mono-testis, which comprises the primer pair;
the primer pair or the kit is applied to the identification of the characteristics of the Duroc pig mono-testis;
the primer pair or the kit is applied to screening of the pigs with the genetic defect of the single testis;
the primer pair or the kit is applied to pig molecular marker assisted breeding;
a method for identifying a molecular marker genotype located on pig chromosome 12 that is associated with formation of pig epididymis, comprising the steps of:
(1) extracting the genome DNA of the pig to be detected;
(2) carrying out PCR amplification on the genomic DNA obtained in the step (1) by adopting the primer pair or the primer pair in the kit to obtain a PCR amplification product;
(3) sequencing the PCR amplification product obtained in the step (2) so as to obtain a sequencing result;
(4) determining the genotype of the molecular marker of the pig to be detected, which is positioned on the pig No. 12 chromosome and is related to the formation of the pig mono-testis based on the sequencing result;
the pig is preferably a pure Duroc and a synthetic line thereof;
a method of genetic improvement in pigs comprising the steps of:
determining the sites of the molecular markers of the pigs in the core pig group, which are positioned on the chromosome 12 of the pigs and are related to the formation of the Duroc monorchidics, and making corresponding selection according to the molecular markers: selecting swine individuals with CC and TC genotypes at 26276230 th position on chromosome 12 of reference sequence of version 11.1 of international swine genome from the swine core group, and eliminating swine individuals with TT genotypes at 26276230 th position to improve the frequency of allele C at the position generation by generation;
the breeding pigs are preferably selected from pure duroc and a synthetic line thereof;
compared with the prior art, the invention has the following advantages and effects:
(1) the invention researches and determines that a molecular marker (the nucleotide mutation with the marked position of the SEQ ID NO:1 sequence as 193) which is obviously related to the formation of the pig single testis character is positioned on the nucleotide sequence on the No. 12 chromosome of the pig, verifies the influence effect of the molecular marker on the single testis character, and applies the molecular marker to the genetic improvement of screening the genetic defect of the pig single testis to ensure the enterprise profit and increase the core competitiveness.
(2) The invention provides a primer pair for identifying a molecular marker which is positioned on a pig No. 12 chromosome and is related to the formation of a pig mono-testis, and provides a new method for screening work related to disease-resistant breeding of pigs by the molecular marker and the primer pair.
Drawings
FIG. 1 is a genome-wide association analysis (GWAS) Manhattan plot of pure Duroc pigs on chromosome 12 for the single-testis trait; wherein: the abscissa represents the chromosome number of the pig; the ordinate represents the-logP value.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
Experimental swinery: in the experiment, 504 pure-breed doloc from a core field of a Swine thigh pig are used together. The swinery can feed and drink water freely, the whole feeding mode, feeding conditions and the like are always consistent, and the method is a conventional method.
Example 1 the process of the invention for obtaining the Gene marker of the invention
(1) The extraction method of pure Duroc pig ear sample tissue DNA refers to the phenol-chloroform labeling method to extract the whole genome DNA. The DNA of the pure Duroc population was subjected to quality detection and concentration measurement using a Nanodrop-ND1000 spectrophotometer. The ratio of A260/280 is 1.8-2.0, and the ratio of A260/230 is 1.7-1.9. Finally, the qualified DNA samples were uniformly diluted to 50 ng/L.
(2) And (3) detecting the 50K SNP genotype of the whole genome of the pig: a GeneSeek Genomic Profile Portine 50K SNP typing platform adopts instructions of Illumina Infinium and standard flow to perform chip hybridization and result scanning. Finally, reading genotype data through genome studio software. Quality control of obtained genotype data by PLINK v1.07, and rejection rate<99.7%, Frequency of the minor allele (Mimor Allel Frequency, MAF)<0.01% or a deviation from Hardy-Weinberg Equilibrium (HWE) P of 10-6SNP marker of (1), exclusion of detection Rate<90%, individuals with a familial mendelian error rate higher than 0.1; the remaining 46462 SNP markers and 504 samples were quality controlled for subsequent data analysis.
(3) Genome Wide Association (GWAS) analysis: in order to eliminate the population stratification effect, the GWAS analysis is carried out by adopting single-point regression analysis of a linear mixed model and combining with an R language GenABEL software package, and the stratification effect is corrected by utilizing the similarity of genomes among individuals in an analysis model. Determining the significance threshold of the SNP and single testis character association degree by adopting a Bonferrini method, wherein the genome level significance threshold is 0.05 divided by the number of effective SNP loci, namely the genome significance threshold is 1.07615 multiplied by 10-6I.e., 0.05/46462 (effective number of SNPs); the chromosome level significance threshold value is 1 divided by the number of effective SNP loci, namely the chromosome significance level threshold value is 2.15230 multiplied by 10-5I.e., 1/46462 (effective SNP number).
The GWAS analysis results are shown in fig. 1. As can be seen from FIG. 1, in Duroc, there is a site in chromosome 12 that significantly affects the pig monorchidism trait, and the most strongly associated SNP is g.193T>C(P＝2.83×10-8)。
(4) Correlation analysis of different genotypes with the porcine testis number phenotype:
as can be seen from Table 1, the SNP site g.193T > C (nucleotide 193 in SEQ NO. 1) of the molecular marker has a very significant association with the characteristics of the mono-testis. In the duroc population, the prevalence of CC genotype pigs is 0.07, the prevalence of TC genotype pigs is 0.26, and the prevalence of TT genotype pigs is 1. Therefore, the TT genotype is preliminarily presumed to be a potential pathogenic genotype for the pig monorchidism character. In addition, the results of the independence test performed based on the genotype frequency indicate that the distribution of the genotype frequency of the SNP site g.193T > C of the molecular marker in the diseased and normal groups is very significantly different (P < 0.01).
TABLE 1 comparison of the distribution of the genotype of SNP site g.193T > C in individuals with single testis disease and normal individuals and allele frequency distribution
Note: the corresponding number of individuals is shown in parentheses; the bolded part is the allele frequency distribution.
Example 2 specifically explains the inventive procedure for detecting SNP marker
(1) The target fragment containing the SNP site obviously related to the Duroc single testis character is a 276bp nucleotide sequence in a chromosome 12, the upstream and downstream primers for sequence amplification are primer-F and primer-R, and the nucleic acid sequences are as follows:
upstream primer-F: 5'-AGAAGCTGGACTTAGAACCC-3';
downstream primer-R: 5'-GAGCCTGGAAAATCTATCCCT-3';
(2) PCR amplification system and condition setting
A10. mu.L system was prepared, in which 1.0. mu.L of DNA sample, 0.3. mu.L of forward primer, 0.3. mu.L of reverse primer, 5.0. mu.L of PCR StarMix, ddH2O3.4. mu.L, PCR conditions of 94 ℃ pre-denaturation for 2min, 94 ℃ denaturation for 30s, 64 ℃ annealing for 30s, 72 ℃ extension for 15s, 35 cycles total, and final extension at 72 ℃ for 5 min.
(3) DNA sequencing identification: the sequence sequencing is carried out in Shenzhen Hua Dagen science and technology Limited, and the gene fragment is used for detecting positive and negative reactions. Comparing the measured sequence with the NCBI genome sequence to obtain the mutation of the corresponding SNP locus, wherein the sequencing result is as follows:
AGAAGCTGGACTTAGAACCCAGGTCTCCTGACTCTAGGCCATGGCCCCATCCAGTCCTCCACCACCTCTTCATCTTGAAGCCAGACCAAAGTATTGGGCCCTGGTTCTCAACCTTGTCCACCTCACAGGTGGGTGGGCATTAATGTAGGGGCAGCCCTACAATGGGCATAAAGAGGTGACACCAGGACCCTGM(T/C)AGCTGCTGTGGTCCCAGCATAGCTCCCCTTAGCCAGGGGCGGGAGGGTGCTCATCTCAAGGTAGGGATAGATTTTCCAGGCTC；
note: m marked in the sequence is a mutation site, indicated by underlining (the mutation base in parentheses, and the allele mutation), and the head and tail of the sequence are shown in bold as the primer sequence binding site.
Example 3 SNP site g.193T > C Effect analysis of molecular markers
As can be seen from Table 1, the SNP site g.193T > C (nucleotide 193 in SEQ NO. 1) of the molecular marker has a very significant association with the characteristics of the mono-testis. In the duroc population, the prevalence of CC genotype pigs is only 0.07, TC genotype pigs is 0.26, and TT genotype pigs is 1. Therefore, by the molecular marker-assisted selection, pigs with the genotype TT in the group are gradually eliminated, the allele frequency of the allele C can be obviously improved, the occurrence rate of the pig monorchidism is reduced, the breeding loss is reduced, and the improvement progress of the pig genetic defect is accelerated, so that the economic benefit of the breeding of the pigs is effectively improved, and the core competitiveness is increased.
The invention provides a new molecular marker for the molecular marker-assisted selection of pigs by detecting the 193 rd base mutation site in the SEQ ID NO.1 sequence and primarily performing the correlation analysis between the genotype and the occurrence of the pig monorchidics, and can establish a high-efficiency and accurate molecular marker-assisted breeding technology by utilizing the molecular marker and a corresponding primer pair.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
SEQUENCE LISTING
<110> southern China university of agriculture; wenshi food group Ltd
<120> molecular marker located on pig No. 12 chromosome and related to formation of pig mono-testis and application
<130> 1
<160> 3
<170> PatentIn version 3.3
<210> 1
<211> 276
<212> DNA
<213> Artificial
<220>
<223> SEQ ID NO：1
<400> 1
agaagctgga cttagaaccc aggtctcctg actctaggcc atggccccat ccagtcctcc 60
accacctctt catcttgaag ccagaccaaa gtattgggcc ctggttctca accttgtcca 120
cctcacaggt gggtgggcat taatgtaggg gcagccctac aatgggcata aagaggtgac 180
accaggaccc tgmagctgct gtggtcccag catagctccc cttagccagg ggcgggaggg 240
tgctcatctc aaggtaggga tagattttcc aggctc 276
<210> 2
<211> 20
<212> DNA
<213> Artificial
<220>
<223> primer-F
<400> 2
<210> 3
<211> 21
<212> DNA
<213> Artificial
<220>
<223> primer-R
<400> 3
gagcctggaa aatctatccc t 21
Claims (7)
1. The application of a detection reagent for detecting molecular markers which are positioned on a pig No. 12 chromosome and are related to the formation of pig mono-testis in the identification of the character and genetic breeding of Duroc pig mono-testis is characterized in that: the SNP locus of the molecular marker which is positioned on the chromosome 12 of the pig and is related to the formation of the pig monocrchidism corresponds to the 26276230 th T > C mutation on the chromosome 12 of the reference sequence version 11.1 of the international pig genome;
the nucleotide sequence of the molecular marker which is positioned on the pig No. 12 chromosome and is related to the formation of the pig mono-testis is shown as SEQ ID NO:1, wherein M in the sequence is T or C, resulting in different numbers of pig testis, and TT genotype is a single testis.
2. A method for detecting the characteristics of the pig monorchidism is characterized by comprising the following steps:
detecting a molecular marker which is located on a pig 12 chromosome and is related to the formation of a pig mono-testis, wherein the molecular marker is located on the pig 12 chromosome according to claim 1, the SNP site mononucleotide of the molecular marker is T or C, and the TT genotype is the mono-testis;
the pig is a pure Duroc and a synthetic line thereof.
3. An application of a primer pair for identifying a molecular marker which is positioned on a pig No. 12 chromosome and is related to the formation of a pig mono-testis in the identification of the character of the Duroc mono-testis is characterized in that:
the molecular marker as set forth in claim 1, wherein the primer pair comprises primer-F and primer-R, and the nucleic acid sequences are as follows:
upstream primer-F: 5'-AGAAGCTGGACTTAGAACCC-3';
downstream primer-R: 5'-GAGCCTGGAAAATCTATCCCT-3'.
4. The application of a primer pair for identifying a molecular marker which is positioned on a pig No. 12 chromosome and is related to the formation of a pig epididymis in screening epididymis genetic defect pigs is characterized in that:
the molecular marker as set forth in claim 1, wherein the primer pair comprises primer-F and primer-R, and the nucleic acid sequences are as follows:
upstream primer-F: 5'-AGAAGCTGGACTTAGAACCC-3';
downstream primer-R: 5'-GAGCCTGGAAAATCTATCCCT-3';
the pig is a pure Duroc and a synthetic line thereof.
5. The application of a kit for identifying a molecular marker which is positioned on a pig No. 12 chromosome and is related to the formation of a pig mono-testis in identifying the character of the Duroc pig mono-testis is characterized in that:
the molecular marker is the molecular marker described in claim 1, and the kit comprises the primer pair described in claim 3.
6. The application of the kit for identifying the molecular marker which is positioned on the chromosome 12 of the pig and is related to the formation of the pig epididymis in screening the pig with the genetic defect of the epididymis is characterized in that:
the molecular marker is the molecular marker described in claim 1, and the kit comprises the primer pair described in claim 3;
the pig is a pure Duroc and a synthetic line thereof.
7. A method of genetic improvement in pigs, comprising the steps of:
determining the sites of the molecular markers related to Duroc mono-testis formation on the chromosome 12 of the pigs in the core group of the pigs according to claim 1, and making corresponding selection according to the molecular markers: selecting swine individuals with CC and TC genotypes at 26276230 th position on chromosome 12 of reference sequence of version 11.1 of international swine genome from the swine core group, and eliminating swine individuals with TT genotypes at 26276230 th position to improve the frequency of allele C at the position generation by generation;
the pig is a pure Duroc and a synthetic line thereof.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010788014.XA CN111850139B (en) | 2020-08-07 | 2020-08-07 | A molecular marker located on porcine chromosome 12 related to the formation of porcine single testis and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010788014.XA CN111850139B (en) | 2020-08-07 | 2020-08-07 | A molecular marker located on porcine chromosome 12 related to the formation of porcine single testis and its application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111850139A CN111850139A (en) | 2020-10-30 |
| CN111850139B true CN111850139B (en) | 2021-12-24 |
Family
ID=72972200
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010788014.XA Active CN111850139B (en) | 2020-08-07 | 2020-08-07 | A molecular marker located on porcine chromosome 12 related to the formation of porcine single testis and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111850139B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105603062A (en) * | 2006-05-03 | 2016-05-25 | 人口诊断股份有限公司 | Method of evaluating genetic disorders |
| CN105779594A (en) * | 2016-03-23 | 2016-07-20 | 西北农林科技大学 | A kind of detection method and application of pig Oct4 gene insertion/deletion |
| CN108849724A (en) * | 2018-06-21 | 2018-11-23 | 安徽浩翔农牧有限公司 | A kind of bar of Du pig and its breeding method |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1581648A2 (en) * | 2002-09-09 | 2005-10-05 | Nura, Inc. | G protein coupled receptors and uses thereof |
-
2020
- 2020-08-07 CN CN202010788014.XA patent/CN111850139B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105603062A (en) * | 2006-05-03 | 2016-05-25 | 人口诊断股份有限公司 | Method of evaluating genetic disorders |
| CN105779594A (en) * | 2016-03-23 | 2016-07-20 | 西北农林科技大学 | A kind of detection method and application of pig Oct4 gene insertion/deletion |
| CN108849724A (en) * | 2018-06-21 | 2018-11-23 | 安徽浩翔农牧有限公司 | A kind of bar of Du pig and its breeding method |
Non-Patent Citations (3)
| Title |
|---|
| Genome-wide association study using deregressed breeding values for cryptorchidism and scrotal/inguinal hernia in two pig lines;Claudia A Sevillano等;《Genetics Selection Evolution》;20150321;第47卷(第1期);第1-8页 * |
| rs336892247;Ensembl;《Ensembl》;20200430;第1页"Alleles"、"Location"部分 * |
| 猪主要遗传缺陷候选基因的研究进展;张蕾等;《猪业科学》;20180825;第35卷(第8期);第116-118页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111850139A (en) | 2020-10-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110117665B (en) | SNP molecular marker located on pig No. 16 chromosome and related to pig lean meat percentage and eye muscle area and application | |
| CN108676897B (en) | SNP marker influencing daily gain traits of pigs and application thereof | |
| CN108315433A (en) | It is a kind of influence Duroc boar intramuscular fat content molecular labeling and application | |
| CN110257529A (en) | SNP marker relevant to lean meat percentage, eye muscle area, eye muscle thickness and application on No. 6 chromosomes of pig | |
| CN105648077B (en) | A kind of molecular marker for affecting daily gain in pigs character and its application | |
| CN110106255B (en) | Molecular markers located on pig chromosome 7 associated with daily weight gain traits in Duroc pigs and their applications | |
| CN110541038A (en) | A SNP molecular marker located on pig chromosome 1 related to pig daily gain and its application | |
| CN108103208A (en) | A kind of SNP marker for influencing sheep Fecundity Trait and its application | |
| CN113564264A (en) | A SNP molecular marker located on pig chromosome 14 and related to the stillbirth number and the healthy litter rate of sows and its application | |
| CN108913779B (en) | SNP marker influencing daily gain traits of pigs and application thereof | |
| CN107267631A (en) | A kind of SNP marker for influenceing daily gain in pigs character and its application | |
| CN107779517B (en) | It is a kind of influence Duroc kind pig lean meat percentage character molecular labeling and its application | |
| CN119320833B (en) | STON1 gene molecular marker primer related to chicken double yolk egg character and application thereof | |
| CN110144408B (en) | Molecular markers and applications of SNPs located on pig chromosome 7 associated with total teat number | |
| CN113736890B (en) | A kind of SNP molecular marker related to the number of healthy piglets and live piglet rate and its application | |
| CN113584185A (en) | SNP molecular marker located on pig chromosome 12 and related to piglet malformation number and application thereof | |
| CN117925850A (en) | SNP locus related to blood heme concentration of blood clam and application of SNP locus in parent breeding | |
| CN111850139B (en) | A molecular marker located on porcine chromosome 12 related to the formation of porcine single testis and its application | |
| CN112280874B (en) | Copy number variation molecular marker influencing pig backfat thickness on pig chromosome 11 and application | |
| CN106701930A (en) | Method for detecting insertion-deletions polymorphism of sheep FTH-1 (ferritin heavy polypeptide1) genes by utilizing PCR-SSCP (polymerase chain reaction-single strand conformation polymorphism) and application thereof | |
| CN112266968B (en) | A molecular marker of copy number variation on chromosome 13 affecting market weight, age and daily gain and its application | |
| CN106755370B (en) | A method for detecting single nucleotide polymorphism of sheep FTH-1 gene by PCR-RFLP and its application | |
| CN112458183B (en) | Copy number variation molecular marker on pig No. 3 chromosome and related to daily gain and weight and age on market of pig and application thereof | |
| CN118957107B (en) | Molecular marker located on chromosome 17 of pig and related to backfat thickness property and application thereof | |
| CN114875028B (en) | Mutation site associated with sow total litter size trait and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |