贵州医科大学学报

2008, No.131(02) 155-158+161

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人幽门螺杆菌HspA亚单位的基因克隆及序列分析
Cloning and Sequence Analysis of Human Helicobacter pylori HspA Subunit Gene

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)

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

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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)

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DOI: 10.19367/j.cnki.1000-2707.2008.02.014

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