鼠疫杆菌F1抗原结构基因caf1在酿酒酵母中的同源重组及初步鉴定

目的研究新型的防治鼠疫的基因疫苗。方法将鼠疫杆菌F1抗原结构基因caf1构建至pHSS6-mTn-3xHA／lacZ转座文库质粒中，将此重组质粒经Not Ⅰ酶切线性化，然后以醋酸锂法转化至酵母细胞中进行同源重组，再以尿嘧啶缺陷型培养基对阳性转化子进行筛选。结果转化子经菌落PCR方法分析鉴定，证明其为重组阳性转化子。结论以同源重组方式构建酿酒酵母表达系统，用以表达鼠疫杆菌F1表面抗原，为经消化道途径实现对鼠疫的基因防治创造条件。

Homologous recombination of caf1 gene of caf operon encoding F1 antigen of Y. pestis in Saccharomyces cerevisiae

Objective To develop a new type of gene vaccine against plague. Methods caf1 gene of caf Operon Encoding F1 Antigen of Y. pestis was inserted into pHSS6-mTn-3xHA/lacZ library plasmids, the recombinant plasmids were linearized with Not I and transformed into yeast cell by lithium acetate (LiAc) method to induce homologous recombination. Positive recombinants were then selected with uracil-lack medium. Results These recombinants were confirmed by colony PCR to have the target gene fragments. Conclusion The present study provided a platform for constructing a novel expression system for expressing Y. pestis F1 Antigen via homologous recombination in Saccharomyces cerevisiae, which may contribute to the genetic prevention of plague through digestive-tract-route (DTR).