Abstract：Abstract: Human cytomegalovirus (HCMV) is commonly found in nature. HCMV gB is an important surface glycoprotein, and its AD1 sequence is one of the major neutralizing epitopes. First, the truncated gB gene containing the AD1 sequence but without the transmembrane and intracellular domains was amplified by PCR using HCMV genomic as template. Then the HCMV gB/AD1 gene was cloned into mammalian cell expression vector pSGHV0, and electroporated with dhfr gene into CHO/dhfr－ cells. Stable cell lines were further screened with 60 nmol/L MTX, and the expression of recombinant AD1 protein was detected by ELISA and Western blot. Finally the fusion protein was purified by Ni2+-NTA affinity chromatography. The recombinant fusion protein was successfully obtained with expression level of about 10 mg/L and purity of over 80%, which can laid solid foundation for further studies of HCMV gB function and genic engineering vaccine research in the future.
引用本文:
董金蓉, 毛树宝, 谢震渊, 沈谊清, 吴金妍, 罗 剑, 方 芳. 人巨细胞病毒gB/AD1基因在CHO细胞中的表达和纯化[J]. 生命科学研究, 2015, 19(5): 402-409. DONG Jin-Rong, MAO Shu-Bao, XIE Zhen-Yuan, SHEN Yi-Qing, WU Jin-Yan, LUO Jian, FANG Fang. Expression and Purification of Human Cytomegalovirus gB/AD1 in CHO Cell. Life Science Research, 2015, 19(5): 402-409.
2015,Vol. 19
