绵羊GDF9基因mRNA、DNA和调控区序列克隆及其在11个品种中遗传多态性检测

doi:doi: 10.11843/j.issn.0366-6964.2016.08.005

Abstract

Abstract:

The aim of this study was to clone the mRNA，DNA and regulatory region sequence of ovine GDF9 gene，detect its genetic polymorphism in 11 sheep breeds，and search for the molecular genetic markers related to sheep litter size.The RACE and PCR technologies were used to clone full-length sequence of ovine GDF9 gene，and SNaPshot was performed to detect the polymorphisms of GDF9 gene in 11 breeds.A 1 852 bp full-length mRNA of ovine GDF9 gene (GenBank No.：KR063137) was obtained，which contains 58 bp 5′-UTR and 432 bp 3′-UTR.In addition，2 898 bp of DNA sequence and 2 304 bp of regulatory region sequence were amplified.Compared with the sequence of NC_019462.1，the result had 2 more fragments in regulatory region.The genetic polymorphism analysis showed that a known mutation G260A (G1) and a novel mutation －2078C>G within regulatory region were identified.The genotyping results showed that 11 sheep breeds were free of FecGE， FecGH， FecGT， FecGF， FecGV mutations，while the G260A and －2078C>G were widespread in all sheep except Prairie Tibetan sheep.G260A contained 3 genotypes AA，BB and AB，but the BB genotype only presented in Small-tail Han sheep and Valley Tibetan sheep.－2078C>G also included 3 genotypes，in which the genotype and allele frequencies were exactly the same with G260A.It seems that －2078C>G was complete linkage with G260A.Association analysis in Small-tail Han sheep showed that the litter size of individuals with AB genotype was significantly higher than that of AA genotype (P＜0.05).We improved the mRNA，DNA and regulatory region sequences of ovine GDF9，which provided a foundation for further functional study of GDF9 gene.Meanwhile，G260A and －2078C>G mutations were found in different sheep breeds，and G260A site could be a potential genetic marker for improving litter size in sheep.

CLC Number:

S826
S821.2

PAN Zhang-yuan，HE Xiao-yun，LIU Qiu-yue，HU Wen-ping，WANG Xiang-yu，GUO Xiao-fei，CAO Xiao-han，DI Ran，CHU Ming-xing. Cloning and Genetic Polymorphism Analysis of mRNA，DNA and Regulatory Region of Ovine GDF9 Gene in 11 Breeds[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(8): 1555-1564.

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Cloning and Genetic Polymorphism Analysis of mRNA，DNA and Regulatory Region of Ovine GDF9 Gene in 11 Breeds

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