鸭乙型肝炎病毒PreS蛋白片段在大肠杆菌中的高效表达及初步应用  

HIGH EXPRESSION OF DUCK HEPATTTIS B VIRUS PRE-S ANTICEN IN E.COLI AND THE APPLICATION OF THIS RECOMBINANT PROTEIN

作　　者：马张妹[1] 李伯良[1] 熊思东[1] 闻玉梅[1] 

机构地区：[1]上海医科大学分子病毒学卫生部重点实验室,中国科学院生物化学研究所

出　　处：《病毒学报》1994年第1期1-7,共7页Chinese Journal of Virology

基　　金：国家863高科技生物领域资助

摘　　要：将编码鸭乙型肝炎病毒（ＤＨＢＶ）前表面抗原（ＰｒｅＳ）第９至１６３位（１５５个）氨基酸序列的４６５ｂｐＤＮＡ片段，克隆入ＰＬ启动子控制下并含有修饰型ｃＩｔｓ８５７基因的表达载体中。利用载体的起始密码子和创建的终止密码子（ＴＡＧ）构建含部分ＤＨＢＶＰｒｅＳ基因的表达质粒。在大肠杆菌ＡＲ６８ｐＲＫ２４８中，本质粒经温敏诱导，表达产物为１７．５ｋＤａ，约占细菌总蛋白的５％。用Ｗｅｓｔｅｍｂｏｌｔ分析，本表达产物可与特异性ＤＨＢＶＰｒｅＳ／Ｓ抗体反应。以表达产物组建固相基质抗体抗原复合物，可部份逆转对ＤＨＢＶ的免疫耐受状态。he DHBV Pre-S fragment encoding amino acid number 9-163 of the Pre-Sprotein,with extra amino acids(Ile Leu)added to its C-terminus and a created stop codon (TAG)was constructed.This fragment was inserted into the pBLMD8 vector.This vectorhas a fixed initiation codon ATG in the PL-controlled transcript and a modified cIts857gene.When the expression vector was transfected tl E.Coli AR68pRK248,and induced at 42℃a product with the expected molecular weight(17.5kDa)was observed,which was approximately 5% of the total bacterial proteins.By Western blatting,the protein expressed retained the reactivity with the specific anti-DHBsAg/Pre-S antibody.The protein expressedwas used to construct Solid-Matrix-Antibody-Antigen Complex(SMAA),which showed some effect to revert the imunotolerance of ducks DHBV.

关 键 词：鸭 乙型肝炎病毒 表达 前S蛋白 

分 类 号：R373.21[医药卫生—病原生物学]