CN105567859A - FSHbeta-gene-based molecular marker related to porcine reproduction traits and detection method thereof, and application of molecular marker - Google Patents
FSHbeta-gene-based molecular marker related to porcine reproduction traits and detection method thereof, and application of molecular marker Download PDFInfo
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- CN105567859A CN105567859A CN201610144670.XA CN201610144670A CN105567859A CN 105567859 A CN105567859 A CN 105567859A CN 201610144670 A CN201610144670 A CN 201610144670A CN 105567859 A CN105567859 A CN 105567859A
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- 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
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- 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
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- C12Q2600/124—Animal traits, i.e. production traits, including athletic performance or the like
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- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Abstract
The invention discloses an FSHbeta-gene-based molecular marker related to porcine reproduction traits and a detection method thereof, and application of the molecular marker. The molecular marker is a porcine FSHbeta gene segment, the porcine FSHbeta gene segment contains a mutant site, and the 8942bp-8954bp position mutated into porcine FSHbeta gene nucleotide sequence has an insertion/deletion of an 11bp base segment. The research detects that the 3rd exon of the porcine FSHbeta gene has a mutant site, and is mutated into an insertion/deletion of the 11bp base. The mutant site is related to porcine reproduction traits, and especially related to total number born or number born alive, and can be easily detected. The invention provides an FSHbeta-gene-based molecular marker related to porcine reproduction traits according to the mutant site, a detection method of the molecular marker and application of the molecular marker in porcine reproduction trait selection, thereby providing bases for porcine molecular breeding and research of the porcine reproduction physiological mechanism.
Description
Technical field
The present invention relates to Animal molecular breeding field, particularly relate to the relevant molecule marker of a kind of pig reproductive trait based on FSH β gene and detection method thereof and application.
Background technology
Molecular marker assisted selection is the important application of genetically engineered in modern cattle breeding, utilize the various mark replacements selection by phenotype based on relevant to breeding character, individual genetic composition is analyzed from molecular level, thus realize genotypic direct selection, the drawback directly selected can be overcome, improve the accuracy of selection and shorten the generation interval.
The reproductive trait of the pig such as total yield coefficient, number born alive is important economic characters, is directly connected to the economic benefit of pig industry.
FSH may grow, maintains spermatogenesis, stimulates interstitial glands to grow and stimulate the follicular development of jenny, promotion endometrial growth by induced male animal convoluted seminiferous tubule in animal HPG Reproductive Axis, also work in coordination with synthesis and the secretion of LH pungency glandular hormone, promote follicle maturity, ovulation.Pig FSH β gene is located in 2p1.6-p1.2, comprises 3 exons and 2 introns.FSH β gene is considered to affect the important candidate gene of of reproductive performance.Association analysis about FSH β gene pleiomorphism and reproductive trait mainly concentrates on people, pig, sheep, Niu Shang.These results prompting FSH β gene may be a major gene controlling reproductive performance, or presents close linkage state with major gene.On pig, an insertion mutation in FSH β gene the 1st intron becomes study hotspot, increasing research display, allelotrope A (insertion mutation) preponderates in place of china pig, and allelotrope B (deletion mutantion) preponderates in external pig, but about the effect of this site to swine reproduction performance, research kind is different, and conclusion is also not quite similar.And the research in other region of FSH β gene is few.
Summary of the invention
Goal of the invention: for problems of the prior art, the invention provides the molecule marker that a kind of pig reproductive trait based on FSH β gene is relevant, molecular breeding for pig provides new thinking, and another object of the present invention is to provide detection method and the application of described molecule marker.
Technical scheme: the molecule marker that the pig reproductive trait based on FSH β gene of the present invention is relevant, it is pig FSH beta gene fragment, described pig FSH beta gene fragment contains mutational site, the insertion/deletion of the base fragment of the 11bp that the 8942bp-8954bp place sporting pig FSH β gene nucleotide series exists.
The nucleotide sequence of pig FSH β gene is as shown in SEQIDNO.4, shown in sudden change SEQIDNO.4, the 8942bp-8954bp of sequence, is in described mutational site, is closely related with pig total yield coefficient, number born alive, be easy to detect, this site can be utilized to develop the molecule marker relevant to pig reproductive trait.
Pig FSH beta gene fragment of the present invention is as shown in SEQIDNO.1.Certainly, pig FSH beta gene fragment of the present invention also can for comprising the DNA fragmentation of the nucleotide sequence as shown in SEQIDNO.1.According to detection method, the DNA fragmentation that can comprise mutational site of appropriate length can be selected.
Present invention also offers the detection method of described molecule marker, comprising:
(1) according to the nucleotide sequence design primer of described molecule marker;
(2) with the genomic dna of pig to be detected for template carries out pcr amplification;
(3) type of pleomorphism site in pcr amplification product is judged.
In step (3), can check order or electrophoretic analysis to PCR primer, judge the base type of pleomorphism site.
Preferably, in step (1), the nucleotide sequence of upstream primer is as shown in SEQIDNO.2, and the nucleotide sequence of downstream primer is as shown in SEQIDNO.3.Utilize this primer directly can carry out pcr amplification, amplified production agarose or polyacrylamide gel electrophoresis carry out genotype tests.
Present invention also offers the application of described molecule marker in pig reproductive trait is selected.
Concrete, pig can be little MeiShan pig, Fengjing pig or Large White.Reproductive trait is total yield coefficient or number born alive.
Compared with prior art, beneficial effect of the present invention is:
The present invention studies discovery, there is a mutational site in pig FSH β gene the 3rd exon, this sports the insertion/deletion of 11bp base, the reproductive trait of this mutational site and pig especially total yield coefficient or number born alive relevant, and easily detect, thus the present invention provides the relevant molecule marker of a kind of pig reproductive trait based on FSH β gene according to this mutational site, and provide the detection method of this molecule marker and the application in pig reproductive trait is selected, for pig molecular breeding, research pig breeding physiology mechanism provide basis.
Accompanying drawing explanation
Fig. 1 is pcr amplification product PAGE gel genotyping result in embodiment 1;
Fig. 2 is GG and HH genotype individuals PCR primer sequencing result in embodiment 1.
Embodiment
Below in conjunction with specific embodiment, illustrate the present invention further.
Embodiment 1
Within 2013, gather the ear sample of 106 little MeiShan pig, 42 Fengjing pig and 70 Large Whites; Arrange the Product archives of above-mentioned 106 the little Mei mountain sows in 2004, Jiangsu Polytechnic College of Agriculture and Forestry's little MeiShan pig breeding center ~ during 2012 simultaneously, had 698 nest piglet records, mainly collect the index such as farrowing time, parity, TNB, NBA.Ear sample (about 0.15g) overbit pincers gather, and-20 DEG C frozen.Extract pig genomic dna with the phenol chloroform extraction method of routine, carry out OD pH-value determination pH, calculate its concentration to genomic dna, and to be diluted to final concentration with ddH2O be 100ng/ μ L, stoste-20 DEG C preservation, diluent 4 DEG C preservation, diluent is as the template of pcr amplification.
The first step: pig FSH β gene complete sequence (accession number: D00621.1) announced according to GenBank, adopt mixed base because of pool technology, amplification pig FSH β gene 3 exons, find after order-checking that in the sequence after pig FSH β gene 8942bp, prompting appears 1-2 peak, in each base positions: the sequence after 8942bp may occur disappearance or insert phenomenon.Wherein, each genes of individuals group DNA sample, because of in pool technology, is drawn 5 μ L and is mixed by mixed base, form DNA pond, as amplification template, the amplimer adopted is: upstream primer F:agcctccaacatcttctt, downstream primer R:attcaatgcctgtctcat.Amplification system, for being 20 μ L, comprising: 10 × Buffer2.0 μ L, 25mMMg
2+2.2 μ L, 10mMdNTPs0.8 μ L, 10 μMs of each 1 μ L of upstream and downstream primer, DNA profiling 1 μ L (100ng), 5U μ L
-1taq enzyme 0.2 μ L, ddH
2o11.8 μ L.Pcr amplification reaction program: 94 DEG C of denaturation 5min; 31 circulations (72 DEG C extend 1min for 94 DEG C of sex change 1min, 62 DEG C of annealing 30s); Last 72 DEG C extend 10min, preserve product for 4 DEG C.
Second step: adopt Primer5.0 software design 1 pair of primer, this base deletion region of increasing.Primer is: FSH β-3del.Upstream primer sequence is: 5 '-atagacccccatctcccaat-3 ', and downstream primer sequence is: 5 '-gactggccctgctcttgtag-3 ', and amplification region is 8917bp-9158bp (SEQIDNO.1), amplified production length 242bp.
3rd step: pcr amplification.Pcr amplification reaction system is 20 μ L, comprising: 10 × Buffer2.0 μ L, 25mMMg
2+2.2 μ L, 10mMdNTPs0.8 μ L, 10 μMs of each 1 μ L of upstream and downstream primer, DNA profiling 1 μ L (100ng), 5U μ L
-1taq enzyme 0.2 μ L, ddH
2o11.8 μ L.Pcr amplification reaction program: 94 DEG C of denaturation 5min; 31 circulations (72 DEG C extend 1min for 94 DEG C of sex change 1min, 60 DEG C of annealing 30s); Last 72 DEG C extend 10min, preserve product for 4 DEG C.Amplified production polyacrylamide gel electrophoresis carries out genotype tests, and genotyping the results are shown in Figure in 1, Fig. 1, and the genotype of swimming lane 3,7 is GG, and the genotype of swimming lane 4,5 is HH, and the genotype of swimming lane 1,2,6,8,9 is GH, M is Marker.
4th step: the pcr amplification product of GG and HH genotype individuals is served Hai Sheng work biotech firm and check order, sequencing result is shown in Fig. 2.
5th step: analyze the distribution situation of this site in different swinery.The results are shown in Table 1, a kind of genotype, i.e. GG in Large White colony, only detected, illustrate that allelotrope G is fixed up in Large White colony; The gene frequency of allelotrope G in little MeiShan pig and Fengjing pig 2 swinerys is respectively 0.580,0.357, χ
2assay shows, 2 swinerys are all in Hardy-Weinberg equilibrium state on FSH β-3.2 site, and (P value is respectively 0.652,0.911; P>0.05).
The genotype frequency in table 13 swinery FSH β gene a 3rd exons 1 1bp base deletion site and allelotype frequency
6th step: SPSS software context analyzes the relation between this mutational site and reproductive trait.The results are shown in Table 2, in 1 ~ 2 tire little Mei mountain sow, the TNB of HH type individuality is 12.09, than GG, GH type individuality high 1.24 (P<0.01), 0.35 (P>0.05) respectively, the NBA of HH type individuality is the highest, be 11.25, but the NBA difference of 3 kinds of genotype individuals is not significantly (P>0.05); In 2 tire above little Mei mountain sows, the TNB of HH type individuality is 13.16, be significantly higher than GG, GH type individuality (P<0.05), not significantly (P>0.05), the NBA difference of 3 kinds of genotype individuals is not all significantly (P>0.05) for the TNB difference of GG and GH type individuality; In the little Mei mountain sow of all parity, the TNB of HH type individuality is significantly higher than GH type individuality (P<0.05), and pole is significantly higher than GG type individuality (P<0.01); The NBA of HH type individuality is significantly higher than GG type individuality (P<0.05), with GH type individual difference remarkable (P>0.05); In 3 stages, TNB and the NBA of 3 kinds of genotype individuals all shows identical trend: HH>GH>GG.
The association analysis of table 2FSH β gene 11 bp-del site and little MeiShan pig reproductive trait
Note: N is nest number; TNB is total yield coefficient; NBA is number born alive
Result is pointed out: on pig FSH β gene 11 bp-del site, the reproductive performance of homozygote HH is better than the reproductive performance of heterozygote and homozygote GG, its additive effect also can be fixed up in more passing from generation to generation, in addition, H gene is lacked in Large White, possible this is that breediness caused, and therefore should strengthen H gene and the genotypic selection of HH in seed selection.
Claims (8)
1. the molecule marker that the pig reproductive trait based on FSH β gene is relevant, it is characterized in that, it is pig FSH beta gene fragment, described pig FSH beta gene fragment contains mutational site, the insertion/deletion of the base fragment of the 11bp that the 8942bp-8954bp place sporting pig FSH β gene nucleotide series exists.
2. molecule marker according to claim 1, is characterized in that, described pig FSH beta gene fragment comprises the nucleotide sequence as shown in SEQIDNO.1.
3. molecule marker according to claim 2, is characterized in that, described pig FSH beta gene fragment is as shown in SEQIDNO.1.
4. a detection method for the molecule marker of being correlated with based on the pig reproductive trait of FSH β gene according to any one of claims 1 to 3, is characterized in that, comprising:
(1) the nucleotide sequence design primer of the molecule marker according to any one of claims 1 to 3;
(2) with the genomic dna of pig to be detected for template carries out pcr amplification;
(3) type of pleomorphism site in pcr amplification product is judged.
5. detection method according to claim 4, is characterized in that, in step (1), the nucleotide sequence of upstream primer is as shown in SEQIDNO.2, and the nucleotide sequence of downstream primer is as shown in SEQIDNO.3.
6. the application of the molecule marker according to any one of claims 1 to 3 in pig reproductive trait is selected.
7. application according to claim 6, is characterized in that, pig is little MeiShan pig, Fengjing pig or Large White.
8. application according to claim 6, is characterized in that, reproductive trait is total yield coefficient or number born alive.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108676898A (en) * | 2018-06-28 | 2018-10-19 | 漳浦县赵木兰养殖有限公司 | With the application of the relevant Gene A NYZ of litter size of pig |
| CN109897903A (en) * | 2019-04-26 | 2019-06-18 | 四川农业大学 | Molecular labeling and application based on FSH β identified for genes Large White reproductive trait |
| CN111996264A (en) * | 2020-09-17 | 2020-11-27 | 湖北省农业科学院畜牧兽医研究所 | Application of pig SNP molecular marker in pig breeding character screening and pig breeding |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102080081A (en) * | 2010-11-19 | 2011-06-01 | 华南农业大学 | SNP (Single Nucleotide Polymorphism) marker related to litter size of sow and application thereof |
| CN102405877A (en) * | 2010-09-20 | 2012-04-11 | 南通天盟养殖有限公司 | Synthetic-line selection-breeding method by utilizing pig mark to assist selection |
| CN105368926A (en) * | 2015-08-27 | 2016-03-02 | 安徽农业大学 | Method for measuring pig gene polymorphism and relevance between pig gene polymorphism and sow reproduction traits |
-
2016
- 2016-03-14 CN CN201610144670.XA patent/CN105567859A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102405877A (en) * | 2010-09-20 | 2012-04-11 | 南通天盟养殖有限公司 | Synthetic-line selection-breeding method by utilizing pig mark to assist selection |
| CN102080081A (en) * | 2010-11-19 | 2011-06-01 | 华南农业大学 | SNP (Single Nucleotide Polymorphism) marker related to litter size of sow and application thereof |
| CN105368926A (en) * | 2015-08-27 | 2016-03-02 | 安徽农业大学 | Method for measuring pig gene polymorphism and relevance between pig gene polymorphism and sow reproduction traits |
Non-Patent Citations (2)
| Title |
|---|
| 吴井生: "猪繁殖性状相关基因遗传效应及表达规律的研究", 《中国博士学位论文全文数据库 农业科技辑》 * |
| 朱孟玲等: "FSHβ基因和PRLR基因的多态性对小梅山猪繁殖性状的影响", 《猪业科学》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108676898A (en) * | 2018-06-28 | 2018-10-19 | 漳浦县赵木兰养殖有限公司 | With the application of the relevant Gene A NYZ of litter size of pig |
| CN109897903A (en) * | 2019-04-26 | 2019-06-18 | 四川农业大学 | Molecular labeling and application based on FSH β identified for genes Large White reproductive trait |
| CN109897903B (en) * | 2019-04-26 | 2022-03-15 | 四川农业大学 | Molecular marker for identifying breeding traits of white pigs based on FSH beta gene and application |
| CN111996264A (en) * | 2020-09-17 | 2020-11-27 | 湖北省农业科学院畜牧兽医研究所 | Application of pig SNP molecular marker in pig breeding character screening and pig breeding |
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