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目的 克隆幽门螺杆菌 (Helicobacterpylori,Hp) 4种黏附素 (babA、alpA、alpB和hopZ)基因的保守区 ,并对其进行序列及生物信息学分析。方法 利用ANTHEPROTV4 .3c软件 ,分析已证实的Hp 4种黏附素蛋白序列 ,发现 4种黏附素的C 端氨基酸存在保守区。选取AlpA的C 端结构基因序列作为引物进行PCR ,将PCR产物定向插入载体pET 2 2b(+) ,以DNA自动分析仪进行序列测定 ,并以生物信息学软件分析其生物学特性。结果 克隆片段的长度为5 88bp ,与发表的黏附素AlpA保守区基因的序列完全一致。ANTHEPROTV4 .3c软件预测其蛋白质的Mr 约为 2 25 0 0 ,并显示出良好的抗原性和疏水性。结论 用PCR方法 ,从幽门螺杆菌ss1中获取到黏附素AlpA保守区的基因序列 ,为研究Hp黏附素的分子机制和免疫原性奠定了基础
Objective To clone the conserved regions of four kinds of adhesins (babA, alpA, alpB and hopZ) genes of Helicobacter pylori (Hp) and analyze their sequence and bioinformatics. Methods Using ANTHEPROTV4.3c software, we analyzed the four protein sequences of Hp and confirmed the existence of C-terminal amino acids of four kinds of adhesins. The sequence of C-terminal AlpA gene was selected as the primer for PCR. The PCR product was inserted into pET 2 2b (+) vector and sequenced by DNA analyzer. Bioinformatics software was used to analyze its biological characteristics. Results The length of the cloned fragment was 5 88 bp, which was completely consistent with the published sequence of AlpA conserved region of adhesin. ANTHEPROTV4 .3c software predicts that its protein Mr is about 2550 and shows good antigenicity and hydrophobicity. Conclusion The sequence of the conserved region of adhesin AlpA was obtained from Helicobacter pylori ss1 by PCR, which laid the foundation for studying the molecular mechanism and immunogenicity of Hp adhesin