大白菜BcpLH基因的原核表达及其蛋白产物的Western分析  

PROKARYOTE EXPRESSION AND WESTERN ANALYSIS OF BcpLH GENE OF CHINESE CABBAGE

作　　者：余旭红[1] 郑志仁[1] 刘平林[1] 何玉科[1] 

机构地区：[1]中国科学院上海植物生理研究所,上海200032

出　　处：《实验生物学报》2000年第4期301-307,共7页Acta Biologiae Experimentalis Sinica

基　　金：国家自然科学基金(项目批准号:39870450)

摘　　要：BcpLH基因是大白菜包叶组织特异的新基因,含有双链RNA结合域。在含有His标记序列的原核表达载体pET28-a(+)上插入Bc-pLH基因的cDNA,在大肠杆菌BL21(DE3)中诱导表达出了特异性蛋白,并免疫大白兔制备出高效价的抗血清;同时,将BcpLH基因插入到含有超级助溶剂的pMAL-c2载体上,并在大肠杆菌DH5α中诱导表达,结果获得了可溶的蛋白。Western斑点印迹分析的结果证明了BcpLH的特异性。BcpLH活性蛋白及其抗血清的产生为研究BcpLH基因的RNA结合活性及其在植物发育上的调节机制创造了条件。BcpLH gene preferentially expressed in folding leaf of Chinese cabbage contains dsRNA-binding domains. The cDNA of BcpLH gene was cloned into a His-fusion expression vector pET-28a (+) and was induced to express in E. colt strain BL21(DE3). Then, the specific protein was partially purified and the rabbit was immunized to prepare the anti-serum. Meanwhile Bc- pLH cDNA was cloned into the pMAL-c2 containing the solubizing partner, and then the soluble protein generated. It was demonstrated from Western dot assay that the BcpLH protein was specific. The BcpLH active protein and its anti-serurn made it possible to study RNA-binding activity and regulation mechanism in plant development.

关 键 词：大白菜 BcpLH基因 原核表达 蛋白产物 

分 类 号：S634[农业科学—蔬菜学]