乙肝病毒表面抗原SA-28融合基因在酵母中的组成型表达  

Expression of the Modified HBsAg Gene SA-28 Directed by a Constitutive Promoter

作　　者：周则迅[1] 袁汉英[2,3] 何炜 高卜渝[2,3] 李育阳[2,3] 

机构地区：[1]复旦大学遗传学研究所,上海200433 [2]复旦大学生命科学学院 [3]复旦大学遗传工程国家重点实验室

出　　处：《复旦学报（自然科学版）》2000年第3期264-268,272,共6页Journal of Fudan University：Natural Science

基　　金：国家科委八六三高科技资助项目 (Z18 0 1)

摘　　要：以酵母载体YFD5 9为基础 ,将乙肝病毒表面抗原SA 2 8融合基因正向插入组成型启动子PPGK1及终止子TPGK1间 ,得到表达载体YFD5 9 LSS1.并将该表达载体分别转化BJ2 16 8,DBY746 ,JRY188,BJ35 0 5 4株不同的酿酒酵母株 .再将YFD5 9 LSS1质粒上的表达单元克隆至高稳定载体pHC11的BamHⅠ位点 .构建成 3个带有不同拷贝数和不同插入方向表达单元的表达载体 :pHC11 LSS1 1,pHC11 LSS1 2和 pHC11 LSS1 7.将它们分别转化Y16 ,Y192株酿酒酵母菌 .对不同工程菌株进行表达水平的测定 ,我们选到了高效表达乙肝病毒融合抗原的菌株 ,并发现带有以YFD5 9为基础的表达载体的工程菌表达水平要比带有以 pHC11为基础的表达载体的工程菌的高 。By placing the modified HBsAg gene SA 28 in a yeast expression vector carrying promoter P PGK1 and terminator T PGK1 , an expression plasmid YFD59 LSS1 was constructed. It was transformed into four yeast strains, BJ2168, DBY746, JPY188, BJ3505. The expression cassette from YFD59 LSS1 was then inserted into the BamHⅠ site of a stable yeast plasmid pHC11. Three expression vectors, pHC11 LSS1 1, pHC11 LSS1 2 and pHC11 LSS1 7, were obtained. They were transformed into two yeast strains, Y16 and Y19. The ELISA assay shown that strains BJ3505/YFD59 LSS1 and DBY746/YFD59 LSS1 could highly express modified HBsAg. The expression level of strains, which carry expression vectors based on plasmid pHC11, is much lower than that of the strains, which carry YFD59 LSS1. The stability of expression vector may be the major reason for the difference of expression level.

关 键 词：乙肝病毒表面抗原 组成型表达 SA-28融合基因 

分 类 号：R392－33[医药卫生—免疫学] Q789[医药卫生—基础医学]