新基因RSTA14-44在大肠杆菌中的表达及抗体制备  

The fusion expression, purification of RSTA14-44 and preparation of its polyclonal antibody

作　　者：袁莉刚[1] 田永强 缪时英 王琳芳 刘英[1] 

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070 [2]中国协和医科大医学分子生物学国家重点试验室,北京100005

出　　处：《甘肃农业大学学报》2004年第5期499-502,共4页Journal of Gansu Agricultural University

基　　金：国家重点基础研究发展规划项目基金(G1999055901)

摘　　要：RSTA14-44 基因是中国协和医科大分子生物学试验室在研究中获得的一个新基因,全长 1124 bp,可读编码框自 100bp^684 bp,编码 194 个氨基酸,预测其编码蛋白质分子量为 21.8 KDa。构建了包含 RSTA14-44 cDNA 全部编码框的 pET30a(+)-14 表达载体质粒,经过聚丙烯酰氨凝胶电泳纯化,免疫家兔制备了抗 RSTA14-44 抗体。Western免疫印迹分析表明,该抗体具有较高的反应性和特异性,适合用作 RSTA14-44 蛋白的进一步研究。RSTA14-44 is a new gene with 1124 bp of full length. It contains a 584 bp of open reading frame, from 100 to 684 bp, encoding a protein with 194 amino acids, and putative protein molecular weight is 21.8 KDa. In present study, the reading fragment of RSTA14-44 gene was cloned into the expression vector pET30a (+). The fusion protein was obtained and purified. Rabbits were immunolized by the fusion protein of RSTA14-44. The titer and specificity of polyclonal antibody were detected by indirect ELISA assay and Western blotting. The results demonstrated the very high specificity of the antibody which could be used for the further study of RSTA14-44 protein.

关 键 词：RSTA14-44 融合表达 多克隆抗体 

分 类 号：Q786[生物学—分子生物学]