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.
出处
《甘肃农业大学学报》
CAS
CSCD
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
