摘要
目的:探讨HCV-core与HCBP1的相互作用.方法:PCR分别扩增含HCV core及HCBP1的cDNA片段,克隆入pGEM T载体,经测序正确后再分别克隆入哺乳双杂交的表达质粒pM和pVP16中,并转染入COS-7细胞,48 h后检测细胞中报告基因CAT的表达水平.结果:CAT-ELISA检测显示pM-core与pVP16-HCBP1实验孔A405 nm值为0.1288,高于其它阴性对照,但低于阳性对照组A值.结论:哺乳双杂交系统证实HCV核心蛋白与新基因HCBP1有一定的相互作用,可进一步研究HCBP1的细胞功能.
AIM： To analyze the interaction between HCV core protein and HCBP1 using mammalian two-hybrid assay. METHODS： cDNA fragments encoding HCV core protein and HCBP1 were amplified by PCR and subsequently cloned into pGEM T vector, respectively. After verified by sequencing, they were respectively subcloned into two hybrid plasmids, pM and pVP16. Then, pM-core, pVP16-HCBP1 and pGSCAT, a reporter vector, all were co-transfected into COS-7 cells. Forty-eight hours later, the interaction between HCV core protein and HCBP1 was assayed by detecting CAT gene expression. RESULTS： CATELISA showed that the A value of the co-transfection experiment group was 0. 1288, significantly higher than those of the other negative control groups and lower than that of the positive control group. CONCLUSION： Mammalian two-hybrid assay confirms HCBP1 can bind with HCV core protein to some extent. Based on this, we can study the function of HCBP1 further.
出处
《第四军医大学学报》
CAS
北大核心
2008年第7期588-591,共4页
Journal of the Fourth Military Medical University
基金
国家自然科学基金(30471532)
陕西省科学技术研究发展计划项目(2004K15-G1)
关键词
HCV核心蛋白
哺乳双杂交
报告基因
hepatitis C virus core protein
mammalian two-hybrid assay
reporter gene
