不同载体-宿主菌组合对重组蛋白表达量的影响  

Effect of Different Vector and Host Combinations on Recombinant Protein Expression

作　　者：杨刚刚[1,2] 王泽 白海静[1,2] 杨亚娟[1,2] 徐存拴 

机构地区：[1]河南师范大学生命科学学院,河南新乡453007 [2]河南省-科技部共建细胞分化国家重点实验室培育基地和河南省生物工程重点实验室,河南新乡453007 [3]河南新乡华星药厂,河南新乡453007

出　　处：《河南科技大学学报（自然科学版）》2016年第2期78-81,87,共5页Journal of Henan University of Science And Technology:Natural Science

基　　金：国家"973"前期研究专项基金项目(2012CB722304);河南省自然科学基金项目(142300413212;132300413208);河南省重大科技攻关基金项目(111100910600)

摘　　要：研究了不同载体-宿主菌组合对毕赤酵母重组蛋白表达量的影响。以豹蛙抗瘤酶(onconase,ONC)、IL-1α、IL-1β、IL-1Ra、人血清白蛋白(human serum albumin,HSA)与ONC的融合蛋白即HSA-(Gly_4Ser_1)_3-ONC(简称HSA-ONC)等5种重组蛋白为报告蛋白,设计并合成了5种基因。分别插入3种酵母表达载体p PIC9、p PIC9K和p PICZα-A中,转化3种毕赤酵母受体菌X-33、GS115和SMD1168。筛选得到7种载体-宿主菌组合,在摇瓶规模进行诱导并定量。研究结果表明:5种外源蛋白在p PICZα-A/X-33组合中表达量最高,在p PIC9/SMD1168组合中最低。从重组蛋白表达量来说,p PICZα-A/X-33组合优于其他6种载体-宿主菌组合。The effect of vector-host combination on the expression level of recombination proteins in Pichia pastoris system was studied. Five recombination proteins including onconase( ONC),IL-1α,IL-1β,IL-1 Ra and HSA-( Gly_4Ser_1)_3-ONC( HSA-ONC) fusion protein were inserted into p PIC9,p PIC9 K and p PICZα-A,then the recombination vectors were respectively transformed into three P. pastoris strains( X-33,GS115 and SMD1168). Seven kinds of vector-host combinations were obtained after being screened,and these five recombination proteins were induced and quantitified in the flask scale. The results show that the expression level of five fusion proteins are the highest in p PICZα-A / X-33,but the lowest in p PIC9 / SMD1168. The p PICZα-A / X-33 system is better than the other six combinations based on the yield of recombination proteins.

关 键 词：载体 宿主菌 载体-宿主菌组合 重组蛋白 表达量 

分 类 号：Q815[生物学—生物工程]