幽门螺杆菌HspA和UreB双价侯选疫苗株的构建

目的 构建表达幽门螺杆菌的组成成分热休克蛋白Ａ亚单位（ＨｓｐＡ）和尿素酶Ｂ亚单位（ＵｒｅＢ）的重组蛋的白质的侯选菌株。方法 用ＰＣＲ方法从幽门螺杆菌的染色休ＤＮＡ上分别扩增出ＨｓｐＡ和ＵｒｅＢ基因片段，将它们融合插入原核表达载体ｐＥＴ－２３ｂ（＋）中，并在ＢＬ２１（ＤＥ３）大肠杆菌表达。结果 经测序，ＨｓｐＡ－ＵｒｅＢ（ＨＵ）事例基因片段由２０６１ｂｐ组成，为编码６８７个氨基酸残基的多肽。ＳＤＳ－

Construction of bivalent vaccine candidate expressing HspA and UreB subunit from Helicobacter pylori

Objective Both heat shock protein A subunit (HspA) and urease B (UreB) subunit as effective immunogen may stimulate the immunoresponse protecting human body against challenge of H.pylori. A recombinant strain which expresses bivalent antigen of HspA and UreB subunit was constructed. Methods Gene encoding the structural A/B subunit of H.pylori heat shock protein/ urease was amplified from H.pylori chromosomal DNA by PCR techniques. The genes were inserted into the prokaryotic expression vector pET 22b( ), and then was transformed into the BL 21(DE3) E.coli strain, expressed which HspA UreB recombinant protein. Results HspA UreB gene was found to be 2061 base pairs ,the recombinant fusion protein encoded polypeptides of 687 amino acid residues, corresponding to calculated molecular masses of 78kD. Western blot analysis of HspA UreB recombinant protein confirmed that it could be specifically recognized by the serum from H.pylori infected patients, and could also be recognized by the serum from BALB/c mice immunized with HspA UreB itself. The urease activity of E.coli cells containing the UreB gene plus the HspA gene was two times greater than that of UreB lonely. Conclusion This result suggested that HspA UreB recombinant protein may be an potential vaccine for control and treatment of H.pylori infection.