日本血吸虫Dna J-类似蛋白基因真核表达载体的构建与鉴定

Construction and identification of recombinant Dna J-like protein of Schistosoma japonicum

目的 构建日本血吸虫Dna J-类似蛋白真核表达载体,以待进行动物实验观察其保护性免疫作用。方法 在已构建的克隆载体p Bluescript SK+ / Sj Dna J-类似蛋白(保存于日本血吸虫成虫c DNA文库中)的基础上,根据Dna J-类似蛋白基因序列设计一对引物,PCR扩增后与T载体连接,T/ A克隆筛选后经Eco R 和Xba 双酶切,连接入真核表达载体pc DNA3.1(+) ,构建重组质粒并转化入DH5α,再进行重组基因鉴定。结果 此重组体转入了完整的日本血吸虫Dna J-类似蛋白基因。结论 构建了日本血吸虫Dna J-类似蛋白真核表达载体。

Objective To construct a Schistosoma japonicum Dna J- like protein eukaryon expression vector. Methods According to the cDNA sequence of S.japonicum Dna J-like protein, a pair of primers was designed. With the two primers, a Sj Dna J-like protein gene was amplified from p Blueseript SK+/Sj Dna J-like protein (in cDNA library of S.japonicum) and con nected with T vector. After di-enzyme- cutting by EcoRⅠ and XbaⅠ, the Sj DnaJ-like protein DNA was subcloned into e ukaryocyte expression vector pcDNA3.1(+). Recombinant plasmids were transformed into DH5α and identified. Results The eukaryo tic plasmids including Sj Dna J-like protein DNA were constructed successfully. Conclusion A S.japonicum Dna J-like protein eukaryon expression vector has been established