免疫筛选日本血吸虫成虫cDNA文库基因的亚克隆与鉴定

Subclone and characterization of clone genes immunoscreened from adult Schistosoma japonicum cDNA library

目的亚克隆免疫筛选日本血吸虫ｃＤＮＡ文库得到的基因。方法在体外将血吸虫ｃＤＮＡ文库基因的ＰＣＲ产物和ｐＧＥＭＴ载体连接，转染大肠杆菌ＸＬ１Ｂｌｕｅ，经抗生素及呈色底物Ｘｇａｌ粗筛，再用限制性内切酶及ＰＣＲ方法进一步鉴定。结果成功地将文库中４个大小不同的日本血吸虫基因片段连接到载体中，获得４个重组子。结论为可能编码保护性抗原的血吸虫基因测序和将其亚克隆入真核表达质粒奠定基础。

Objective To construct clone of genes encoding candidate vaccine antigens. Methods Adult S japonicum cDNA library was screened and some positive clones were amplified by PCR. PCR products were ligated into pGEM T vector and then used to transform E coli XL1 blue. Following blue and white selection screening, restriction enzymes and PCR were used to characterize the recombinants. Results Four different pieces of S japonicum cDNA genes were cloned to pGEM T vector. Conclusion Construction of those clones will facilitate further to subclone the genes encoding S japonicum protective antigens into eukaryotic expressive plasmid and to determine their sequence.