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SNP marking primers related to Erhualian living piglet number born and application thereof
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A technology of Erhualian pigs and Erhualian, which is applied in the field of molecular biology, can solve the problems that the main effect gene and its key variation sites cannot be determined, and the genetic mechanism of high fecundity has not been fully revealed and effectively utilized. The effect of rapid detection
Active Publication Date: 2019-02-19
NANJING AGRICULTURAL UNIVERSITY
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[0003] Erhualian pig is a high-quality pig breed in the Taihu Lake Basin in my country. It is famous all over the world for its high litter size and is an important resource for the sustainable development of my country's pig industry; although a large number of domestic research institutions have used Erhualian to identify this pig breed The genetic mechanism of high fecundity, but limited by many factors such as research methods, means, materials and the complexity of reproductive traits, the genetic mechanism of high fecundity of the Erhualian pig breed has not been fully revealed and effectively utilized
[0005] According to the International Porcine QTL Database website (http: / / www.animalgenome.org / cgi-bin / QTLdb / SS / index), there is currently no location on chromosomes 10, 11 and sex chromosomes in pigs that affects the total litter size. QTLs affecting total litter size have been located on other autosomes, but most of them are QTLs located by microsatellite markers, and the confidence interval is mostly 10-20cM, so it is impossible to determine the real main effectgene and its key variation Therefore, it is difficult to be directly applied to the breeding improvement of breeding pigs, so finding the molecular marker sites that affect the litter size is the key to increase the litter size
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Embodiment 1
[0028] 1. Source of experimental animals
[0029] Jiaoxi Erhualian pig professional cooperative in Changzhou, Jiangsu.
[0030] Calculate the breeding value of 177 Erhualian sows, the calculation model is
[0031] Y (total litter size) = parity (parity) + farm (field) + year (annual) + season (season) + age (sow farrowing age) + sire (matingboar) + permanent effect (female Pig's permanent effect) + additive effect (individual additive effect),
[0032] These included fixed effects - parity, farm / year / season sows farrowed, covariates - age of sows farrowed, random effects - boar with mating, permanent effects - sows, individual additive genetic values. 25 individuals with higher breeding value and 25 individuals with lower breeding value were selected.
[0034] Ear tissue samples from 48 sows were collected, placed in centrifuge tubes filled with 75% alcohol, and stored in a -20°C refrigerator for later use.
[0035] Using the traditiona...
Embodiment 2
[0059] In this example, SNPs (SNP / 16kb) were screened for Erhualian pigs within 500 kb upstream and downstream of the GWAS candidate sites obtained in Example 1 to obtain fine-mapped sites.
[0060] 1. Extraction of Genomic DNA from Erhualian sows
[0061] 266 ear tissue samples of purebred Erhualian sows with accurate live birth records were collected and placed in centrifuge tubes filled with 75% alcohol, and stored in a -20°C refrigerator for later use. Genomic DNA from ear tissue was extracted by the above method, and after quality and concentration detection, the concentration was diluted to 30 ng / μL and stored at -20°C for future use.
[0062] 2. Fine positioning of the target area
[0063] The DNA of the above-mentioned individuals was sent to Shanghai Tianhao Biotechnology Co., Ltd. for fine positioning, and many SNPs including the g.80866737 site were obtained, and then SAS software was used to analyze the effect of genotype on phenotype on these SNPs. Analysis mode...
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Abstract
The invention discloses SNP marking primers related to the Erhualian living piglet number born and application thereof. An SNP marking site is located on a nucleotide sequence of a calcium isolinelingene CLSTN2 on a pig 13th chromosome, is a g.80866737 nucleotide site on the pig 13th chromosome according to an international pig genome 11.1-version reference sequence and has C / T polymorphism; theSNP marking is remarkably related to the living piglet number born of Erhualian female pigs. A primer pair for detecting the SNP marking includes the upstream primer shown in SEQ ID NO:2 and the downstream primer shown in SEQ ID NO:3. The SNP marking is related to the living piglet number born of the Erhualian female pigs, and therefore the high-yield Erhualian pig species can be screened out by authenticating the SNP marking, and the obtained Erhualian pig high-yield species has great economic benefits and social value.
Description
technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a SNP marker primer related to the live-born traits of Erhualian pigs and an application thereof. Background technique [0002] In the breeding of commercial pig lines, reproductive traits such as litter size and teat number are particularly important in the selection of dam lines. As one of the important production traits of sows, litter size reflects the level of sow fecundity, pig farm production level and economic effect. With the development of society and economy, people's living standards generally improve, and the demand for pork is also increasing. According to data from the National Bureau of Statistics and Zhue.com, the number of live pigs in my country in 2017 was 433.25 million, and the number of slaughtered pigs was 688.61 million. my country's pig stock, slaughter and meat production account for about half of the world. The level of litter size is direct...
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