德保矮马生长激素受体基因PCR-SSCP分析

Analysis of growth hormone receptor gene of Debao pony by PCR-SSCP

为了研究德保矮马生长激素受体(GHR)基因的多态性,试验根据GenBank公布的马GHR基因样序列设计了9对特异性引物,PCR扩增出德保矮马GHR基因的9个外显子片段(eoxn 2～10),单链构象多态性(SSCP)分析这些片段多态性,并进行群体遗传学分析。结果表明:德保矮马GHR基因9个片段中有3个片段表现出聚合酶链式反应-单链构象多态性,即E1、E3和E5,其他6个片段无多态性。E1片段表现为3种基因型AA、GG和AG型,GG型为优势基因型,G为优势等位基因;E3片段表现出4种基因型,即AA、AC、GA和GC,GA为优势基因型,A、G为优势等位基因;E5片段表现出3种基因型,即AA、AG和GG型,GG为优势基因型,G为优势等位基因。GHR基因E1、E3、E5片段均为中度多态;E1和E3片段处于Hardy-Weinberg非平衡状态,E5处于平衡状态。

To study the polymorphism of growth hormone receptor （GHR） gene of Debao pony, nine pairs of specific primers were designed according to the horse growth hormone receptor gene - like sequences published in GenBank, nine fragments from exon 2 to exon 10 of GHR gene of Debao ponies were amplified by PCR, and the polymorphism of these fragments was analyzed by single - strand conformation polymorphism （SSCP） and population genetics analysis. The results showed that there were three of nine fragments showing polymerase chain reaction - single strand conformation polymorphism （ PCR - SSCP）, including E1, E3 and ES, and the others six fragments showing non - polymorphism. There were three genotypes in E1 fragment, namely AA, GG and AG, in which GG type was the dominant genotype, and G was the dominant allele. There were four genotypes in E3 fragment, namely AA, AC, GA and GC, in which GA was the dominant genotype, both A and G were the dominant alleles. There were three genotypes in E5 fragment, namely AA, GG and AG, in which GG was the dominant genotype, and G was the dominant allele. It was found that E1, E3 and E5 fragments of GI-IR gene were in the moderate polymorphism. The Hardy - Weinberg equilibrium test indicated that E1 and E3 fragments were in a non- equilibrium state, but E5 fragment was in a state of equilibrium.