分泌型乙型肝炎病毒包膜M蛋白在毕赤酵母中的表达  

Expression of the Secretory Hepatitis B Virus Middle Surface Protein in Pichia pastoris

作　　者：丁云菲[1] 刘勇[1] 刘红岩[1] 马雁冰[1] 孙强明[1] 孙茂盛[1] 戴长柏[1] 

机构地区：[1]中国协和医科大学中国医学科学院医学生物学研究所分子生物室,昆明650118

出　　处：《中国生物化学与分子生物学报》2002年第3期265-271,共7页Chinese Journal of Biochemistry and Molecular Biology

摘　　要：将乙肝病毒包膜中蛋白M基因插入酵母整合型表达质粒pAO818的醇氧化酶 (AOX1)启动子下游 ,构建携带 8拷贝M表达盒的重组载体 ,经电穿孔转化SMD116 8菌株和G4 18筛选 ,得到了高效分泌表达M蛋白的毕赤酵母菌株 ,表达量超过 5 0mg L .经初步纯化 ,对表达产物的性质鉴定表明 ,重组蛋白具有preS2和S抗原性 ,可以形成颗粒 ,并具有一定程度的糖基化 .所构建的稳定重组菌株和所得到的重组蛋白颗粒 ,为进一步研究新一代的疫苗提供了必要的材料 .The methylotrophic yeast Pichia pastoris has been developed as production systems for recombinant proteins recently. The favourable and most advantageous characteristics of the species have resulted in an increasing number of biotechnological applications. Gene of hepatitis B virus middle surface protein (M) was placed under the control of AOX 1 promoter in an integrative Pichia expression vector carrying eight copies of the expression cassette. Through electroporation transformation of SMD1168 and G418 selection, a strain of Pichia pastoris capable of secreting M protein into the medium at a level up to 50 mg/L was constructed. After initially purified, the characterization of the expression products demonstrated that the recombinant protein could be assembled into particles which presented preS2 and S antigenicity and a certain extent of glycosylation. These results have provided a good foundation for further research and development of new hepatitis B virus vaccine.

关 键 词：分泌型乙型肝炎病毒 包膜M蛋白 毕赤酵母中 表达 高拷贝重组质粒 包膜中蛋白 分泌表达 基因工程疫苗 

分 类 号：R512.62[医药卫生—内科学] R373.21[医药卫生—临床医学]