论文部分内容阅读
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 ,这可能与相应的表达载体在酵母细胞中的稳定性高低有关
Based on the yeast vector YFD5 9, the SA 2 8 fusion gene of hepatitis B virus surface was inserted between the constitutive promoter PPGK1 and the terminator TPGK1 to obtain the expression vector YFD5 9 LSS 1. The expression vector was transformed into BJ2 16 8, DBY746, JRY188 and BJ35 0 5, respectively.And then the expression unit on the YS-1 vector of YFD5 9 was cloned into the BamHⅠ site of the high-stable vector pHC11.The constructed expression vector was constructed with 3 different expression vectors with different copy number and insertion direction The expression vectors of pHC11 LSS1 1, pHC11 LSS1 2 and pHC11 LSS1 were transformed into Saccharomyces cerevisiae strains Y16 and Y192.According to the expression levels of different engineering strains, we selected the strains highly expressing hepatitis B virus fusion antigen, It was found that the expression level of engineered bacteria with YFD5 9-based expression vector was higher than that of engineered bacteria with pHC11-based expression vector, which may be related to the stability of the corresponding expression vector in yeast cells