摘要
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pHSS6mTn3xHA/lacZ转座文库质粒中,将此重组质粒经NotI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。
Objective To develop a new type of gene vaccine against plague. Methods call 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）.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2005年第9期822-824,共3页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金资助项目(批准号:39880044)
关键词
鼠疫杆菌
F1抗原
酿酒酵母
同源重组
Y. pestis
F1 antigen Saccharomyces cerevisiae Homologous recombination
