构建人Delta-like4ext-26-217原核表达载体并表达.采用PCR方法扩增hDll4ext-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pET32a(+)原核表达载体,获得pET32a(+ )-hDll4ext-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、Western Blot方法鉴定目的蛋白的表达.结果表明,采用SDS-PAGE,Western Blot等方法均可检测到目的蛋白TRX/hDll4ext-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/hD114ext-26-217 fusion protein prokaryotic expression vector, the cDNA sequence of hDl14ext-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/hD114ext-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.
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