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幽门螺杆菌基因hp0231和hp0410的克隆表达及生物信息学分析
引用本文:胡平,罗军,张文炳,龙北国.幽门螺杆菌基因hp0231和hp0410的克隆表达及生物信息学分析[J].热带医学杂志,2007,7(4):319-322,349.
作者姓名:胡平  罗军  张文炳  龙北国
作者单位:南方医科大学公共卫生与热带医学学院微生物教研室,广州,510515
基金项目:广东省广州市科技攻关项目
摘    要:目的获取幽门螺杆菌hp0231和hp0410基因的序列,预测其编码蛋白HP0231和HP0410作为候选疫苗的可行性,在大肠杆菌表达系统中表达hp0231和hp0410。方法提取幽门螺杆菌标准株NCTC11639的基因组DNA,按照GenBank中幽门螺杆菌标准株22695的序列设计引物PCR,扩增hp0231和hp0410基因,克隆入pMD18-T载体中测序,进行生物信息学分析。将hp0231和hp0410克隆入表达载体pGEX-4T-1中,诱导表达,SDS-PAGE电泳鉴定。结果生物信息学分析表明HP0231和HP0410均为外膜蛋白,具良好的抗原性,并且与其他生物的同源性较低,其作为Hp疫苗不易产生交叉反应以及自身免疫性反应,构建的重组菌可高水平表达可溶性重组蛋白。结论HP0231和HP0410是有良好应用前景的Hp候选疫苗。

关 键 词:hp0231和hp0410基因  幽门螺杆菌  克隆表达  生物信息学分析
文章编号:1672-3619(2007)04-0319-04
修稿时间:2006-11-202007-01-27

Cloning, Expression and Bioinformatics Analysis of Helicobacter pylori Genes hp0231 and hp0410
HU Ping,LUO Jun,ZHANG Wen-bin,LONG Bei-guo.Cloning, Expression and Bioinformatics Analysis of Helicobacter pylori Genes hp0231 and hp0410[J].Journal Of Tropical Medicine,2007,7(4):319-322,349.
Authors:HU Ping  LUO Jun  ZHANG Wen-bin  LONG Bei-guo
Institution:School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China
Abstract:Objective To analyze the features and predict the vaccination potential of the proteins endcoded by hp0231 and hp0410 genes,and to express the proteins in E.coli.Method Specific primers of Helicobacter pylori(Hp)gene hp0231 and hp0410 were designed according to the DNA sequence of the Hp strain 22695 released in GenBank of NCBI.These primers were used to amplify the hp0231 and hp0410 from the Hp NCTC11639 genomic DNA by PCR.The PCR fragment was cloned into pMD18-T and then transformed into E.coli TOP10.The sequences of these two genes were analyzed by bioinformatics softwares.The hp0231 and hp0410 gene were then cloned into pGEX-4T-1 and successfully expressed in E.coli.Result The predicted amino acid sequences of the two genes showed that the two proteins are out-membrane proteins and have significant antigenicity.The sequences also showed a low homology with other species so that it is unlikely that the antibodies against these two proteins will cross-react with the corresponding proteins of other species in the immunoassay,and will not cause self immune response.The recombination bacterias could express soluble proteins at high level.Conclusion Our work suggests that HP0231 and HP0410 are potential vaccine candidates of Hp.
Keywords:hp0231 and hp0410  Helicobacter pylori  clone and express  bioinfromatic analysis
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