摘要
化学合成血小板源性生长因子受体结合域１３肽基因，并与乙肝病毒核心抗原基因５′端融合，序列分析表明化学合成的１３肽基因及融合后基因的阅读框架正确．将融合基因亚克隆于ｔａｃ启动子控制的ｐＥＴ３ａ表达质粒中并于大肠杆菌中表达．表达产物经ＥＬＩＳＡ、ＷｅｓｔｒｅｎＢｌｏｔ鉴定表明，融合蛋白已被表达，其单位分子量与推算值一致．电镜观察证明所表达的融合蛋白能形成颗粒．
In order to block the effect of the platelet derived growth factor (PDGF) in atherosclerosis,the fusion gene of PDGF receptor binding domain was cloned with Hepatitis B virus core antigen(HBcAg).The fusion protein was expressed in E.coli and purified by chromatography.The expressed protein served as an immunogen to immunize the rats.The effect of immunogen in genesis of atherosclerosis was obaerved.The DNA fragment encoding PDGF receptor binding domain,ANFLVWEIVRKKP,a 13 mer peptide,was synthesized and fused with the Hepatitis B Core Antigen (HBcAg)gene.The fusion gene was analyzed by DNA sequencing and expressed in E.coli under the control of the tac promoter.The products were analyzed by ELISA and Western Blot,which confirmed that the hybrid protein was expressed as expected.The protein was seen to form particles under electron microscope.The result showed the rats immunized with the fusion protein produced two antibodies,13 mer peptide and HBcAg.The immunogen of PDGF binding domain was produced sucessfully.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
1998年第5期525-530,共6页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金
关键词
乙肝病毒
核心抗原
融合蛋白
PDGF
Platelet derived growth factor,Hepatitis B,Core antigen,Fusion protein
