重组人肿瘤坏死因子α和白细胞介素6融合基因的构建和表达

Construction and expression of recombinant human tumor necrosis factor and interleukin 6 fusion gene

本文利用PCR技术和基因定位突变技术 ,将编码人肿瘤坏死因子α(hTNFα)和白细胞介素 6 (hIL 6 )成熟肽的基因通过中间接头连接成编码单一蛋白的基因 ,构建了融合蛋白表达载体 pBIT ,并在大肠杆菌中得到了表达。SDS PAGE的电泳胶薄层扫描显示 ,融合蛋白的表达量是菌体总蛋白量的 2 0 % ,其分子量约为 37kD。活性检测证实 ,融合蛋白既有TNF活性 ,又有IL 6活性。

An expression vector,in which human tumor necrosis factor alpha(hTNFα)and interleukin 6(IL 6)gene were connected to code a hybird protein by a linker sequence,was constructed by means of PCR amplification and site specific mutagenesis.The fused gene was expressed in E.coli DH5 α and the scanning of SDS PAGE gel showed that the amount of the expressed fussion protein is about 20% of total cell proteins.The recominant fusion protein,which has a molecular weight of estimated 37 kDa,showed both TNF and IL 6 biological activity.