真核表达载体pcDNA3.1(+)-MyoD-HA-His的构建及功能检测  

Construction and effect test of the eukaryotic expression vector pcDNA3.1(+)-MyoD-HA-His

作　　者：杜朝[1] 玉蕾叶 孙国杰[1] 王英泽[1] 

机构地区：[1]河北科技大学生物科学与工程学院,河北石家庄050018

出　　处：《河北科技大学学报》2014年第2期149-153,共5页Journal of Hebei University of Science and Technology

基　　金：国家自然科学基金(81171455);国家自然科学基金青年科学基金(31100715);河北科技大学博士科研启动基金(自然科学)

摘　　要：为了便于检测和纯化转染到非肌肉细胞的生肌因子MyoD,同时为能够与转染的其他生肌因子的表达量进行比较,将MyoD编码区克隆在真核表达载体pcDNA3.1(+)-HA-His。测序表明克隆的MyoD序列正确,并与标签序列构成一个开放阅读框;Western blot显示在起始密码子前添加kozak序列GCCGCCACC,可使融合蛋白能够以较高水平表达;生肌转化实验表明,融合蛋白MyoD-HA能够有效激活靶基因的表达。In order to conveniently detect and purify the myogenic factor MyoD exoexpressed in the nonmuscle cells, and to compare with the expression level of other myogenic factors cotransfected with MyoD, the coding sequence （CDS） of MyoD was cloned and inserted into pcDNA3. I（＋）HAHis, an eukarytic expression vector. Sequencing result confirms that the in sert is correct and forms the same ORF with the HA and its sequences. Furthermore, Western blot shows that the fusion pro tein MyoDHA expresses with a high level for the addition of kozak sequence GCCGCCACC,and myogenic conversion demon strates the fusion protein MyoDHA can activate the target genes effectively.

关 键 词：表达载体 标签 MYOD 

分 类 号：Q781[生物学—分子生物学]