摘要
利用ｐＧＥＭ－Ｔ载体对ＣｐｐＢ基因ＰＣＲ扩增产物进行直接克隆，除常规方法中的限制性内切酶可以节省外，发现该方法不仅过程简便，操作易行，而且可以用ＰＣＲ方法筛选重组子，克隆的阳性率较常规方法高。ＤＮＡ序列测定结果证明在ＰＣＲ产物３′末端的确出现了一个不与模板互补的“Ａ”末端，它可以与ＰＧＥＭ－Ｔ载体Ｔ结合。ＰＣＲ方法筛选重组，克隆的阳性率较常规方法高。ＤＮＡ序列测定结果证明在ＰＣＲ产物克隆片段的基因序列与文献报道相一致，这为获得淋球菌隐蔽性质粒ＣｐｐＢ基因ＰＣＲ诊断标准阳性模板，了解ＣｐｐＢ基因功能奠定了基础。
Directly cloning CppB gene PCR products of the cryptic plasmid of neisseria gonorrhoeae into pGEM-T vector can not only save the restriction endonucleases used in the ordinary cloning method, but also process simply and operate conveniently. The recombination plsmids can be selected with PCR method and the recombination positive rate is found higher than the ordinary method. DNA sequencing results proves that there is an addtion of an adenosine at the 3 end of the PCR products which can combine with the thymidine of the pGEMT vector. The DNA sequene obtained is the same as the published one before. All the above is helpful to obtain the standard template of PCR diagnosis kit of CppB gene of neisseria gonorrhoeae and to understand the function of CppB gene of the cryptic plasmid.
关键词
淋球菌
CppB基因
克隆
序列测定
Neisseria gonorrhoeae CppB gene Cloning DNA sequencing
