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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 R基因片段
In order to construct a bivalent oral live vaccine candidate strain of Vibrio cholerae and Leishmania donovani, the authors amplified and cloned tox R, a virulence-regulated expression gene of V. cholerae by PCR, and restricted the tox R gene of V. cholerae Digestion analysis. The results showed that all of the seven strains of V. cholerae amplified a 1.3 kb fragment of tox R gene. The tox R gene contained the Eco R restriction site in the middle and the tox R gene was cloned into the plasmid p AT1 5 3. Restriction endonuclease analysis of the recombinant plasmid pt R4 confirmed that the DNA fragment inserted into the plasmid pt R4 was a tox R gene fragment