近交五指山小型猪SLA-DQA基因的克隆及在大肠杆菌的原核表达  

Cloning and prokaryotic expression of SLA-DQA gene of inbred miniature Wuzhishan pig

作　　者：王晓东[1] 牟玉莲[2] 张莉[2] 杨树林[2] 罗玉柱[1] 冯书堂[2] 

机构地区：[1]甘肃农业大学动物科学技术学院,甘肃兰州730070 [2]中国农业科学院畜牧研究所,北京100094

出　　处：《甘肃农业大学学报》2006年第4期23-26,共4页Journal of Gansu Agricultural University

基　　金：由科技部"863"(2003AA205100)课题;科技基础性工作(2001DEA10006);社会公益研究专项(2003DEB6J078)课题共同资助

摘　　要：采用RT-PCR方法扩增五指山猪SLA-DQA基因cDNA,克隆入测序载体,测序结果与Gene Bank(登陆号:AY243104)的序列相同,大小为768 bp,该基因编码255个氨基酸残基和一个终止密码,预计蛋白分子量为28 kD.将这个基因克隆到原核表达载体PET-28a(+)中,经酶切、PCR扩增和测序分析确证其正确,插入到表达载体中,阅读框是正确的,从而构建成重组表达载体PET-28a(+)-DQA.重组质粒转化至宿主菌BL21(DE3)中,用IPTG进行诱导表达,对表达的蛋白用SDS-PAGE电泳分析,结果表明得到了与预计分子量相同的蛋白.The eDNA of SLA-DQA gene of inbred miniature Wuzhishan pig was amplified by RT- PCR,and was cloned into PGEM-T vector for sequencing and analysis. The sequence of SLA-DQA cDNA showed no difference with the known sequence in the Gene bank. The SLA-DQA eDNA was cloned into expression vector PET--8a（＋）, after enzyme digestion and sequence analysis, it was testified that the open reading frame was right, then the recombinant expression vector PET-28a（q-）/SLA-DQA was constructed. The PET-28a （＋）/SLA-DQA was transformed into host bacterium, BL21 （DE3）, and was induced to express by IPTG, the purpose protein were identified by SDS-PAGE. The results indicated that the target proteins expressed by Escherichia coli were the same as expected.

关 键 词：SLA—DQA基因 CDNA 克隆 原核表达 

分 类 号：Q78[生物学—分子生物学]