摘要
目的:构建并鉴定pMDl8-T-ipaH重组质粒载体,为探索采用二聚体蝎形探针荧光更精确的定量检测志贺菌标准品的方法奠定基础。方法:以志贺菌袭性相关位的ipaH基因为模板,采用PCR进行扩增,电泳回收纯化,经酶切后克隆入载体质粒pMDl8-T中,最终构建重组质粒pMDl8-T-ipaH,进行酶切和测序鉴定。结果:成功克隆出151 bp左右的ipaH基因片段,导入质粒后,经检酶切及测序鉴定质粒基因片段与目标基因序列一致。结论:成功的构建了pMDl8-T-ipaH重组质粒,为进一步检测志贺菌建立标准品奠定了前期基础。
Objective：To construct and identify the recombinant vector of ipaH gene,which will be used to establish a novel realtime fluorescent PCR method to detect the ipaH gene of Shigella with high sensitivity,specificity and stability using the duplex scorpion probe.Methods： Based on the ipaH gene sequence of strain M32063 of Shigella flexneri recorded in the GenBank,ipaH cDNA was obtained by using PCR and cloned into pMD18-T.At last,pMDl8-T-ipaH was identified by enzyme and sequencing identification.Results： The sequence of ipaH cDNA obtained from the recombinant vector was identical with that published on GeneBank.Conclusion： The recombinant vector pMDl8-T-ipaH was successfully constructed,which lay the foundation for further study.
出处
《中国卫生检验杂志》
北大核心
2013年第5期1183-1185,共3页
Chinese Journal of Health Laboratory Technology
基金
江苏出入境检验检疫局科研计划项目(2012KJ45)
关键词
质粒
ipaH
构建
鉴定
Plasmid
ipaH
Construction
Identification
