吴升伟;吴升宇;苑天红;王明永;吴承龙;
WU Shengwei1,WU Shengyu2,YUAN Tianhong1,WANG Mingyong3,WU Chenglong1(1.Department of Microbiology,Guiyang Medical College,Guiyang 550004,Guizhou,China;2.Caotang Township Centre Hospital of Weng'an County,Weng'an 550400,Guizhou,China;3.Department of Microbiology and Immunology,Xinxiang Medical College,Xinxiang 453003,Henan,China)

• 1:贵阳医学院微生物学教研室
• 2:瓮安县草塘镇中心医院
• 3:贵阳医学院微生物学教研室
• 4:新乡医学院微生物与免疫学教研室

摘要(Abstract)：

目的:获取幽门螺杆菌(Hp)热休克蛋白A(HspA)亚单位基因并进行核苷酸序列分析和同源性比较。方法:获取幽门螺杆菌的染色体DNA,PCR技术扩增获取HspA基因,将其定向插入pGEX-4T-1原核表达载体中进行核苷酸序列分析。结果:经酶切鉴定及基因序列证实HpHspA基因克隆成功,DNA序列与Genbank公布的序列比较有13个碱基存在差异,同源性为96%;由碱基序列推定编码的氨基酸残基序列没有发生变异,同源性为100%。结论:成功地将HpHspA基因克隆到了pGEX4T-1原核表达载体,为HspA的表达和研究奠定了实验基础。
Objective:To get heat shock protein A(HspA)subunit gene of Helicobacter pylori,analyze its DNA sequence,and compare its homology.Methods:Chromosome DNA of H.pylori was extracted and the HspA gene was amplified with Polymerase Chain Reaction(PCR).The obtained DNA fragment was cloned into prokaryotic expression vector pGEX-4T-1 and sequenced.Results:HspA gene was cloned successfully which was demonstrated by enzyme digesting and gene sequencing methods.Comparing with DNA sequences reported in Genbank,nucleotide sequence of the obtained DNA fragment was 96% identical with that of HspA gene with a variation of 13 base pairs.The homology in putative amino acids was 100%.Conclusions:HspA gene has been Cloned into prokaryotic expression vector pGEX 4T-1 successfully,which makes an experimental foundation for its expression and associated research.

关键词(KeyWords)： 螺杆菌,幽门;HspA亚单位;克隆,生物;基因顺序
Helicobacter pylori;HspA subunit;cloning,organism;gene order

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 吴升伟;吴升宇;苑天红;王明永;吴承龙;
WU Shengwei1,WU Shengyu2,YUAN Tianhong1,WANG Mingyong3,WU Chenglong1(1.Department of Microbiology,Guiyang Medical College,Guiyang 550004,Guizhou,China;2.Caotang Township Centre Hospital of Weng'an County,Weng'an 550400,Guizhou,China;3.Department of Microbiology and Immunology,Xinxiang Medical College,Xinxiang 453003,Henan,China)

Email:

DOI: 10.19367/j.cnki.1000-2707.2008.02.014

参考文献(References)：

• [1]Parsonnet J,Friedman GD,Vandersteen DP,et al.Heli-cobacter pyloriinfection and the risk of gastric carcinoma[J].N Engl J Med,1991(325):1127-1131.
• [2]Gomez Duarte OG,Lucas B,Yan ZX,et al Protection of mice against gastric colonization byHelicobacter pyloriby single oral dose immunization with attenuated Salmonella typhimurium producing urease subunits A and B[J].Vaccine,1998(5):460-471.
• [3]Ferrero RL,Thiberge JM,Kausau I,et al.The GroES homolog ofHelicobacter pyloriconfers protective immunity against mucosal infection in mice[J].Proc Natl Acad Sci USA,1995(14):6499-6503.
• [4]Marchetti M,Arico B,Burroni D,et al.Development ofa mouse model ofHelicobacter pyloriinfection that mimics human disease[J].Science,1995(5204):1655-1658.
• [5]黄琛,佘菲菲,陈月秀.幽门螺杆菌CagA蛋白肽段的基因克隆、表达及抗原性检测[J].福建医科大学学报,2003(37):60-63.
• [6]范学工,夏华向.幽门螺杆菌感染——基础与临床[M].长沙:湖南科学技术出版社,1997:33-35.
• [7]Kansau I,Guillain F,Thiberge JM,et al.Nickel binding and immunological properties of the C-terminal domain of thehelicobacter pyloriGroES homologue(HspA)[J].Mol Microbiol,1996(5):1013-1023.
• [8]Suerbaum S,Thiberge JM,Kansau I,et al.Helicobacter pylorihspA-hspB heat-shock gene cluster:nucleotide se-quence,expression,putative function and immunogenicity[J].Mol Microbiol,1994(5):959-974.
• [9]Langenberg W,Rauws EA,Widjojokusomo A,et al.I-dentification of Campylobacter pyloridis isolates by restric-tion endonuclease DNA analysis[J].J Clin Microbiol,1986(3):414-417.

文章评论(Comment)：