乙型肝炎病毒PreS_2S基因在毕赤酵母中表达的研究  

Expression of HBV PreS_2S Gene in Pichia pastoris

作　　者：李利 金立杰[1] 杨屹[1] 邵丽军[1] 谷丽娟[1] 刘岩松[1] 董厉子[1] 王宇[1] 

机构地区：[1]长春生物制品研究所,吉林长春130062

出　　处：《微生物学杂志》2006年第5期44-48,共5页Journal of Microbiology

摘　　要：构建重组质粒PHIL-D2/PreS2S以研究乙肝病毒PreS2(120-146)S基因编码蛋白在毕赤酵母中的表达。通过PCR扩增获得PreS2S片段,插入含AOX1启动子的Pichia Pastoris表达载体PHIL-D2中,构建重组表达质粒PHIL-D2/PreS2S,转化酵母宿主菌GS115。挑取阳性克隆摇床培养,甲醇诱导表达。通过ELISA、RPHA鉴定表达产物。成功构建了PHIL-D2/PreS2S真核表达载体,经过序列分析,插入的基因为在中国流行的adr亚型。在毕赤酵母中重组载体表达了S蛋白,S蛋白的表达量为34.9 mg/L,PreS2抗原检测为强阳性。利用毕赤酵母表达系统能够有效地表达乙型肝炎病毒的PreS2S蛋白,PreS2S蛋白具有良好的生物学活性。In order to construct recombinant plasmid PHIL-D2/PreS2S and express HBV PresS2S gene in Pichia pastoris, the PreS2S gene amplified by PCR from PMD-18T/S2S was inserted into the P. pastoris expression vector PHIL-D2 containing AOX1 promoter. The recombinant expression plasmid PHIL-D2/PreS2S was constructed and transformed into GS115. The positive transformants were induced by methanol for the expression of the PreS2S gene. Products after the induction were analyzed with ELISA and RPHA. The results showed that recombinant plasmid of PHIL-D2/PreS2S has been constructed successfully and sequence analysis suggested that the inserted foreign gene belong to adr subtype, which is the epidemic type in China. The PreS2S gene has been expressed successfully in P. pastoris. Expression quantity of S protein is 34.5 mg/L and PreS2 antigen is strongly positive. Therefore, PreS2S gene has been expressed successfully with P. pastoris vector. And the PreS2S protein in the test has good bioactivity.

关 键 词：PreS2基因 S基因 毕赤酵母 表达 

分 类 号：R373.21[医药卫生—病原生物学]