细粒棘球蚴抗原B酵母表达载体的克隆与序列分析(英文)  

CLONING AND SEQUENCING OF YEAST EXPRESSION VECTOR FOR ECHINOCOCCUS GRANULOSIS ANTIGEN B*

作　　者：卢晓梅[1] 张利华 张文宝 林仁勇[1] Philip Craig McManus Don 温浩[1] 

机构地区：[1]新疆医科大学第一附属医院包虫病临床研究所,乌鲁木齐830054 [2]澳大利亚昆士兰医学研究中心 [3]英国桑福德大学生物科学系

出　　处：《中国寄生虫病防治杂志》2003年第3期159-161,共3页Chinese Journal of Parasitic Disease Control

基　　金：国家自然科学基金项目 (No.39860 0 78);新疆维吾尔自治区自然科学基金项目 (No.990 1 0 30 0 3)。~~

摘　　要：目的　获得重组细粒棘球蚴抗原 B(Eg B)的真核表达蛋白 ,构建 Eg B酵母表达重组体 ,并对构建的重组体进行序列分析。方法　以原核重组体 JM10 9- p Mal2 x Eg B作为模板 ,采用聚合酶链式反应 (PCR)扩增 Eg B片断。目的基因 Eg B片断插入酵母表达载体 p YES2 / NT B,再将酵母表达重组体转化 DH5 a菌 ,并对转化筛选的重组体进行序列分析。　结果　共筛选得到 16个重组克隆 ,其中 7个克隆证实为 Eg B正确序列 ,并与酵母表达载体的开放阅读框 (ORF)相一致。　结论　成功地将细粒棘球蚴抗原 B基因构建入酵母表达载体 p YES2 / NT B的开放阅读框。此重组体可在酵母中进一步表达特异性蛋白。Objective To obtain yeast expression recombination for Echinococcus granulosis antigen B (EgB) and analyze the recombinant yeast vector. Methods EgB fragment as template was extracted from prokaryotic recombinant(JM109-pMal2xEgB) and amplified by PCR. The target gene was ligated into yeast expression vector pYES2/NT B, then the recombination was transformed into DH5α lines and sequenced. Results 16 recombinant colonies were obtained, and 7 contained the correct EgB sequence and were suitable to open reading frame (ORF) of yeast expression vector. Conclusion The recombinant EgB was successfully constructed into yeast expression vector, and it's sequence showed the same result as Shepherd reported. The correct insertion was confirmed within yeast expression vector pYES2/NT B open reading frame by sequencing. The recombination EgB can be used for the expression of specific protein in yeast.

关 键 词：细粒棘球蚴 抗原B 酵母表达载体 克隆 序列 聚合酶链式反应 

分 类 号：R383.33[医药卫生—医学寄生虫学]