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对一株临床分离的高粘附力幽门螺杆菌 (Helicobacterpylori ,Hp)菌株M 1进行粘附素基因hpaA的序列分析 ,为研究Hp的粘附提供分子基础。 方法 :设计hpaA的特异引物 ,将PCR产物插入pUC19载体构建hpaA的重组克隆 ,DNA自动分析仪进行序列测定 ,ClustalX和Homology软件分析DNA及其推导出的氨基酸序列。 结果 :构建了粘附素HpaA的重组质粒 pUC hpa ,测序显示hpaA结构基因长 783bp ,开放读框完整 ,无中断 ,与文献报道的序列有差异 ;该基因编码 2 61个氨基酸 ,其中一段氨基酸序列KRTIQK与大肠杆菌粘附素K99、SfaS、CFA/Ⅰ中的涎酸结合位点的结构相似。结论 :Hp粘附素基因hpaA在不同菌株存在差异 ,但其粘附作用有相似的分子基础。
Sequence analysis of the adhesin gene hpaA of a clinically isolated high adhesion Helicobacter pylori (Hp) strain M 1 provided the molecular basis for the study of Hp adhesion. Methods: Specific primers for hpaA were designed. PCR products were inserted into pUC19 vector to construct recombinant clones of hpaA. The DNA was analyzed by automatic analyzer. ClustalX and Homology software were used to analyze DNA and deduced amino acid sequence. Results: The recombinant plasmid pUC hpa was constructed, and the hpaA gene was 783bp in length. The open reading frame was complete and uninterrupted, which was different from those reported in the literature. The gene encoded 2161 amino acids, of which an amino acid sequence KRTIQK is similar in structure to the sialic acid binding site in E. coli adhesins K99, SfaS, CFA / I. CONCLUSION: The hpaA gene of Hp adhesin is different in different strains, but its adhesion has a similar molecular basis.