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克尔曼尼羊GDF9基因外显子2点突变的鉴定

Identification of point mutations in exon 2 of GDF9 gene in Kermani sheep.

作者信息

Khodabakhshzadeh R, Mohammadabadi M R, Esmailizadeh A K, Moradi Shahrebabak H, Bordbar F, Ansari Namin S

出版信息

Pol J Vet Sci. 2016;19(2):281-9. doi: 10.1515/pjvs-2016-0035.

Abstract

Screening the fertile ewes from national herds to detect the major genes for prolificacy is an effective way to create the fertile flocks. Growth differentiation factor (GDF) 9 is a member of the transforming growth factor β superfamily that is essential for folliculogenesis and female fertility. The aim of this study was to detect single nucleotide polymorphisms (SNPs) in exon 2 of GDF9 gene in Kermani sheep breed using PCR-SSCP. Genomic DNA was extracted from whole blood of collected samples using salting-out method. Whole exon 2 of GDF9 gene was amplified (634 bp and 647 bp fragments) using designed specific primers. The single stranded conformation polymorphism (SSCP) patterns of PCR products were studied using electrophoresis on acrylamide gel and silver-nitrate staining method. Finally, 4 banding patterns for the first primer pair and 4 banding patterns for the second primer pair were obtained. Also, indices of population genetic per SNP were calculated using Gen Alex 6.41 software. The sequencing results showed the presence of 3 mutations (SNP) (443, 477 and 721 positions) in the studied population.

摘要

从国家畜群中筛选能育母羊以检测多产主基因是培育能育羊群的有效方法。生长分化因子(GDF)9是转化生长因子β超家族的成员,对卵泡发生和雌性生育能力至关重要。本研究的目的是利用PCR-SSCP检测克尔马尼绵羊品种GDF9基因外显子2中的单核苷酸多态性(SNP)。采用盐析法从采集样本的全血中提取基因组DNA。使用设计的特异性引物扩增GDF9基因的整个外显子2(634 bp和647 bp片段)。采用丙烯酰胺凝胶电泳和硝酸银染色法研究PCR产物的单链构象多态性(SSCP)模式。最后,第一对引物获得了4种条带模式,第二对引物获得了4种条带模式。此外,使用Gen Alex 6.41软件计算每个SNP的群体遗传指标。测序结果表明,在所研究群体中存在3个突变(SNP)(第443、477和721位)。

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