安徽医科大学学报2012，Vol.47Issue(6)：617-622,6.
基孔肯雅病毒包膜蛋白E2的全基因合成及原核表达
Gene synthesis,cloning,prokaryotic expression and analysis of chikungunya virus E2 glycoprotein
摘要
Abstract
Objective To analyze the impact of hydrophobic transmembrane domain of chikimgunya vims E2 glyco-proteins on E2 expression in E. Coli by comparing E2 expression with the hydrophobic transmembrane domain-deleted mutant. Methods On-line software ExPasy was used to predict transmembrane domain of E2 protein and optimized gene sequence encoding E2 protein was obtained according to amino acids of the protein from GenBank; Primers were designed to synthesize E2 gene according to the principle of OE-PCR and prokaryotic expression plasmids of full-length E2 protein and its mutant were constructed. After being transfromed into E. Coli BL21( DE3 ), two plasmids in E. Coli were induced to express by IPTG and protein expression was identified by SDS-PAGE. Results In the study, the gene encoding CHIKV-E2( 1 - 404 aa ) protein was synthesized by OE-PCR and two prokaryotic expression plasmids,and pET21b-E2( 1 -404 aa ) and pET21b-E2( 1 -350 aa ) were constructed successfully. The plasmids were induced by IPTG and SDS-PAGE, which showed that the production of the deletion mutant E2( 1 -350 aa ) was much higher than that of pET21b-E2( 1 -404 ). Conclusion The hydrophobic transmembrane domain ( 351 -378 aa ) of E2 protein plays an important role in E2 production in E. Coli and production of the mutant with the domain deleted is significantly higher than that of full-length E2 protein.关键词
CHIKV-E2/跨膜疏水性蛋白/全基因合成/缺失突变体/原核表达Key words
CHIKV-E2/ hydrophobic transmembrane protein/ gene synthesis/ truncated mutants/ prokaryotic expression分类
医药卫生引用本文复制引用
章萍萍,邓松华,潘卫,曹洁,陈秋莉,王锦红,张华群,葛宜兵,祁培培,刘超..基孔肯雅病毒包膜蛋白E2的全基因合成及原核表达[J].安徽医科大学学报,2012,47(6):617-622,6.基金项目
安徽省教育厅自然科学重点项目(编号:KJ2010A177) （编号:KJ2010A177）
安徽省自然科学基金(编号:090413136) （编号:090413136）
