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单增李斯特菌新疆临床分离株毒力基因的克隆和序列分析
引用本文:于佳佳,邓兴梅,蒋建军,朱江虹,马勋.单增李斯特菌新疆临床分离株毒力基因的克隆和序列分析[J].石河子大学学报,2014(2):153-157.
作者姓名:于佳佳  邓兴梅  蒋建军  朱江虹  马勋
作者单位:石河子大学动物科技学院预防兽医学重点实验室,石河子832003
基金项目:国家自然科学基金项目(31160506),石河子大学SRP项目(20130188),家畜疫病病原生物学国家重点实验室开放基金课题(SKLVEB2011KFKT007)
摘    要:为了研究单增李斯特菌新疆羊源临床分离的4b血清型菌株(简称LM90)毒力基因的变异情况,参考Genbank上收录的单增李斯特菌4b血清型菌株F2365(GeneBank登陆号AE017262)的Ami、Auto、InlB、InlC、InlJ基因序列,分别设计引物,对LM90的毒力基因Ami、Auto、InlB、InlC、InlJ的最大开放阅读框进行PCR扩增,回收基因序列,连接到PMD19-T载体上进行克隆,筛选阳性菌进行序列的测定,将序列提交到NCBI上(登录号分别为KF826613、KF826614、KF826611、KF703749、KF826612),测序后用DNAMAN软件对序列进行分析。结果显示:LM90的毒力基因Ami、Auto、InlB、InlC、InlJ与F2365毒力基因Ami、Auto、InlB、InlC、InlJ的同源性分别为98.04%、96.53%、98.20%、96.41%、98.23%,氨基酸的同源性分别为99.73%、99.83%、98.67%、99.66%、99.76%,可见单增李斯特菌新疆临床分离株LM90毒力基因Ami、Auto、InlB,InlC、InlJ序列相对稳定。

关 键 词:单增李斯特菌  毒力基因  克隆

Cloning and Analysis of Virulence Genes of Listerisa Monocytogenes in Xinjiang Clinical Strains
YU Jiajia,DENG Xingmei,JIANG Jianjun,ZHU Jianghong,MA Xun.Cloning and Analysis of Virulence Genes of Listerisa Monocytogenes in Xinjiang Clinical Strains[J].Journal of Shihezi University(Natural Science),2014(2):153-157.
Authors:YU Jiajia  DENG Xingmei  JIANG Jianjun  ZHU Jianghong  MA Xun
Institution:(Key Laboratory of Preventive Veterinary Medicine,Animal Science and Technology,Shihezi University,Shihezi 832003,China)
Abstract:The aim of this study was to investigate the variation of virulence genes (Ami,Auto,InlB,InlC and InlJ) of Listeria monocytogenes serotype 4b LM90 isolated from clinical diseased sheep in Xinjiang.The open reading frames (ORFs) of five genes were amplified by PCR with specific primers designed according to the nucleotide sequence of serotype 4b F2365 (accession number: AE017262).The PCR products were purified and cloned into pMD19-T vector,and then positive colonies were screened and sequenced.These gene sequences of Ami,Auto,InlC and InlJ were submitted to GenBank (accession numbers:KF826613,KF826614,KF826611,KF703749,and KF826612,respectively) and analyzed by DNAMAN.Alignment results indicated that the homology of the five virulence genes was 98.04%,96.53%,98.20%,96.41%,and 98.23% at nucleotide sequence level,and 99.73%,99.83%,98.67%,99.66% or 99.76% at amino acid sequence level between LM90 and F2365, respectively.Our research results revealed that the five tested virulence genes of Listeria monocytogenes LM90 are relatively stable.
Keywords:Listerisa monocytogenes  virulence genes  cloning
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