摘要
为了构建霍乱弧菌和杜氏利什曼原虫双价口服活疫苗候选株 ,作者采用 PCR对霍乱弧菌毒力表达调控基因 tox R进行扩增及克隆 ,并对霍乱弧菌 tox R基因进行限制性酶切分析。结果显示 :7株霍乱弧菌均扩增出 1 .3kb的 tox R基因片段 ,tox R基因中部含有 Eco R 酶切位点 ,再将 tox R基因克隆在质粒 p AT1 5 3上 ,对所获的重组质粒 pt R4进行限制性酶切分析 ,证实质粒 pt R4插入的 1 .3kb的 DNA片段为 tox
In order to construct a genomic bivalent oral vaccine of Leishmania donovani and Vibrio cholerae, we amplified a 1.3kb DNA fragment from 7 strains of Vibrio cholerae with primers P1 and P2. Restriction endonuclease analysis of PCR amplified products from 9 strains of Vibrio cholerae was performed by digestion with EcoRⅠ. The results revealed an EcoRⅠ site in the central part of toxR gene. The entire toxR gene of Vibrio cholerae Non CT strain 7743 was amplified by PCR with primers P1 and P2, digested with endonucleases BamHⅠ and Hind Ⅲ, then was orientationally cloned into plasmid pAT153. The restriction endonuclease analysis demonstrates that the recombinant plasmid ptR4 contains a 1.3kb toxR gene fragment.
出处
《华西医科大学学报》
CAS
CSCD
2000年第1期27-29,共3页
Journal of West China University of Medical Sciences
基金
国家自然科学基金! (批准号 3 9870 65 6)
华西医科大学科研基金! ( L 1180 )资助
关键词
霍乱弧菌
毒力表达
调控基因
toxR基因
Vibrio cholerae Virulence expression regulatory gene Gene amplification ToxR gene
