人Delta-like4^(ext)-26-217蛋白原核表达载体的构建及表达  

Construction and Expression of Human Delta-like4^(ext)-26-217 Fusion Protein Prokaryotic Expression Vector

作　　者：黄斯勇[1] 伍艳兰[2] 刘利[1] 李国辉[1] 尹郸丹[1] 何飞[3] 张萍[3] 韩骅[3] 梁英民[1] 

机构地区：[1]第四军医大学唐都医院血液科,西安710038 [2]第四军医大学唐都医院传染科,西安710038 [3]第四军医大学基础部医学遗传学与发育生物学教研室,西安710033

出　　处：《科学技术与工程》2011年第22期5254-5257,共4页Science Technology and Engineering

基　　金：国家自然科学基金面上项目(30871090)资助

摘　　要：构建人Delta-like4ext-26-217原核表达载体并表达。采用PCR方法扩增hD ll4ext-26-217多核苷酸序列,克隆入pMD18T载体。测序正确后,将其亚克隆入pET32 a(+)原核表达载体,获得pET32 a(+)-hD ll4ext-26-217载体。以该载体转化E.coli菌株BL21,IPTG诱导其表达。采用SDS-PAGE、W estern B lot方法鉴定目的蛋白的表达。结果表明,采用SDS-PAGE,W estern B lot等方法均可检测到目的蛋白TRX/hD ll4ext-26-217,分子量约42.2 KD。主要以包涵体形式大量存在。以上结果为Detla-like4功能的进一步研究奠定了基础。To Construction and expression of human TRX/hDll4^ext-26-217 fusion protein prokaryotic expression vector, the cDNA sequence of hDll4^ext-26-217 was amplified by PCR, and cloned into the vector pMD18T. After sequencing, the correct fragment was cloned into prokaryotic expression plasmid pET32a （ ＋ ）. The target protein was expressed in E. coli BL21 induced by IPTG and the expression was detected by SDS-PAGE and Western blotting. The fusion protein TRX/hDll4^ext-26-217 was detected by SDS-PAGE and Western blotting, and mainly aggregated into the inclusion body, which will be useful for further research on the function of Detla-like4.

关 键 词：Detla—like4 NOTCH信号途径 原核表达 血管发生 

分 类 号：Q513.1[生物学—生物化学]