猪肌肉素基因的cDNA克隆与表达  

Cloning and Expression of the Pig Skeletal Muscle Musclin Gene

作　　者：王伟杰[1] 李宏基[1] 韩立强[1] 王月影[1] 王静[1] 李卫华[1] 林茂旺[1] 台玉磊[1] 张志强[1] 藏猛[1] 王艳龄[1] 杨国宇[1] 

机构地区：[1]河南农业大学动物生理生化实验室,郑州450002

出　　处：《生物工程学报》2008年第7期1248-1252,共5页Chinese Journal of Biotechnology

基　　金：河南省重点科技攻关项目(No.0523010500)资助~~

摘　　要：从人肌肉素基因出发，在dbEST数据库中进行同源性搜索，找到七个有较高同源性的Expressed Sequence Tag（DY426490，CF787546，AJ660979，AJ664670，AJ663820，AJ680159，DN106254）。通过拼接和进一步RT-PCR实验验证，获得猪肌肉素基因全长cDNA序列，其全长651bp，开放阅读框为54～452bp，编码有132个氨基酸。同源性分析结果表明，与人、小鼠和大鼠的肌肉素基因cDNA编码区（CDS）同源性分别为87．2％、77．6％和77．9％。利用克隆出的猪肌肉素cDNA，构建表达载体pGEX-4T-1-musclin，并在BL21大肠杆菌中成功表达和纯化了分子量为38．59kD的融合蛋白GST-Musclin，并运用蛋白印迹技术进行鉴定。We found seven tag sequence with high homology in dbEST by using human musclin gene, and got its cDNA sequence, which consists of 65 lbp and the open reading frame was 54-452 bp detected by RT-PCR, encoding 132 amino acid residue protein. The new gene has high homology with that of human, mouse and rat, the rate is 87.2%, 77.6% and 77.9%, respectively; the gene fragment was cloned into expression vector pGEX-4T-1, and the recombinant was transformed into E. coli BL21. Induced by IPTG, the fusion protein GST-musclin, a 38.59 kD protein was successfully expressed in E. coli BL21 and identified by Western blotting.

关 键 词：猪 肌肉素基因 表达 纯化 鉴定 

分 类 号：Q78[生物学—分子生物学]