TAT-NPY融合蛋白的表达和生物活性检测  

Expression and Bioactive of TAT-NPY Fusion Protein

作　　者：费东亮[1] 张才[2,3] 刘国文[2] 王哲[2] 

机构地区：[1]辽宁医学院畜牧兽医学院,辽宁锦州121000 [2]吉林大学,长春130021 [3]河南科技大学,河南洛阳471003

出　　处：《中国农学通报》2012年第17期101-104,共4页Chinese Agricultural Science Bulletin

基　　金：教育部高等学校博士点基金项目资助"NPY对围产期奶牛干物质摄入的调控作用"(20070183156);辽宁省教育厅资助项目"重组神经肽Y对奶牛摄食调控作用影响的研究"(2009B121)

摘　　要：为了获得重组TAT-NPY融合蛋白,并研究其生物学活性。利用基因工程技术,从犊牛下丘脑组织中克隆得到神经肽Y基因序列,通过基因比对符合率为100%。同时,在牛神经肽Y基因序列中插入TAT蛋白基因序列,构建原核表达载体pGEX-3X-NPY-PTD,导入到BL21宿主菌中,进行原核表达,并对表达产物进行鉴定、表达条件优化和纯化,并且经过活性检测,融合蛋白对鸡胚血管生成起促进作用。结果表明:含有PTD结构的重组奶牛神经肽Y具有生物功能活性,为下一步的研究提供了物质基础。In order to obtain the recombinant protein NPY containning TAT protein transduction domain and study the bloactive of the recombinant protein. NPYgene was cloned from the Calf Hypothalamus tissue by the molecular biology techniques. Coincidence rate was 100% by genetic test. TAT-protein gene was inserted into bovine NPY gene sequences and expression vector pGEX-3X-NPY-TAT was constructed, then the recombinant plasmids of pGEX-3X-NPY-PTD was induced in E. coli DE3 by addition of IPTG. The expressed fusion protein was identified, It was optimized and the bioactive test was carried out by CAM. The results showed that the bovine NPY gene of TAT protein transduction domain was expressed, and the expressed fusion protein had biological activity. Conclusion the study provided important base for the animal experiment.

关 键 词：奶牛神经肽Y TAT蛋白 表达 检测 生物活性 

分 类 号：S856.5[农业科学—临床兽医学]