幽门螺杆菌中性粒细胞激活蛋白DNA疫苗的构建及鉴定

目的 构建幽门螺杆菌(Hp)中性粒细胞激活蛋白(HP-NAP)DNA疫苗。方法 扩增全长napA(编码HP-NAP)，将其与克隆载体pBT连接，经测序及同源性分析后，将相应片段亚克隆入真核表达载体pIRES，经双酶切及PCR鉴定。结果 PCR扩增-435bp产物，核苷酸序列测定及BLAST分析表明，所克隆序列与基因库中Hp悉尼株(SS1)napA核苷酸及蛋白质的同源性均为98％。ANTHEPROT软件预测其蛋白质具有良好的疏水性和抗原性。经PCR及双酶切鉴定，证实成功构建携带，napA的重组真核表达质粒plRES-napA。结论 成功构建并鉴定了HP-NAP重组DNA疫苗，为进一步研究其免疫原性及研制口服DNA疫苗奠定了基础。

Construction and identification of a recombinant DNA vaccine of Helicobacter pylori neutrophil activating protein

Objective To construct a recombinant DNA vaccine of Helicobacter pylori (H. pylori) neutrophil activating protein (HP NAP). Methods The 435bp napA gene (encoding HP NAP) was amplified by polymerase chain reaction (PCR) and cloned into TA cloning vector pBT. After nucleotide sequencing and BLAST analysis, the target sequence was then subcloned into a eukaryotic expression vector pIRES and followed by identification of restriction endonuclease digestion and PCR.Results A recombinant eukaryotic expression plasmid pIRES napA containing HP NAP gene of H. pylori was successfully constructed. Most of the H. pylori napA sequence was homologized with that of SS1 strain provided by GenBank, and the homology of neucleotide and protein was 98%. ANTHEPROT software predicted that it possessed good hydrophobicity and antigenicity. Conclusions The successful construction and identification of a recombinant DNA vaccine of HP NAP may help to study its immunogenicity and develop multivalent oral nucleotide vaccine against H. pylori infection.