【摘要】 为研究中国特色品系荷包猪SLA-DRa基因(又称SLA-DRa-HB),本试验设计引物,RT-PCR扩增3个个体荷包猪SLA-DRa全基因编码区,并克隆至pMD18-T载体,转化大肠杆菌JM109感受态细胞,经酶切鉴定后筛选阳性克隆测序,比较分析与其他SLA-DRa等位基因的差异,并绘制分子进化树。结果显示,RT-PCR成功扩增出目的基因条带,大小约800bp。经克隆测序后分析,SLA-DRa-HB基因全长为779bp,编码区为1—759,共编码252个氨基酸。序列对比分析结果显示,SLA-DRa-HB的特征性变异集中在135、159、202位点。而穿膜区和胞浆功能区(203—252)变异位点为206、248。分子进化树分析显示,SLA-DRa-HB自成一系,且与其他等位基因的进化关系较近。本研究成功克隆荷包猪SLA-DRa基因,为进一步研究其功能奠定基础。更多还原

【Abstract】 In order to study SLA-DRa gene(also named as SLA-DRa-HB)from one of special breed of domestic pig,Hebao,primers were designed to amplify the coding domain of SLA-DRa from three Hebao pigs.Then,the amplified fragments were cloned into pMD18-T vector followed by transforming them into Escherichia coli JM109.After cleaving by the restricted enzymes,the positive clones were selected to be sequenced.The differences between SLA-DRa-HB and other SLA-DRa alleles were analyzed by sequences comparing,and then the phylogenetic tree was constructed.The results showed that the interest of the fragments was successfully amplified by RTPCR and the molecular weight was about 800 bp.By cloning and sequencing,the whole length of SLA-DRa-HB was 779 bp and the open reading fragments(ORF)of SLA-DRa-HB located at sites of 1to 759 coding for 252 amino acids.Results of sequences comparision and analysis showed that the characterized amino acids of SLA-DRa-HB mainly focused on sites of 135,159 and 202,while the mutational sites in trans-membrane and cytoplasm were 206 and 248.Analyzing from the phylogenetic tree,it showed that SLA-DRa-HB was clustered into one branch and they were relatively closed to other SLA-DRa alleles.In this research,the SLA-DRa alleles were cloned successfully from Hebao pigs and it would lay a base for study the function of the SLA-DRa genes.更多还原