细粒棘球蚴抗原B酵母表达载体的克隆与序列分析

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。

CLONING AND SEQUENCING OF YEAST EXPRESSION VECTORFOR ECHINOCOCCUS GRANULOSIS ANTIGEN B

Objective To obtain yeast expression recombination for Echinococcus granulosis antigen B (EgB) and analyze the recombinant yeast vector.MethodsEgB 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.Results16 recombinant colonies were obtained,and 7 contained the correct EgB sequence and were suitable to open reading frame (ORF) of yeast expression vector.ConclusionThe 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.