幽门螺杆菌26kDa外膜蛋白的基因克隆

目的:构建幽门螺杆菌外膜蛋白的基因重组载体,为的疫苗开发、快速诊断试剂盒的研究奠定基础。26kDaHp方法:用从染色体基因组扩增外膜蛋白的基因片段,将目的基因、载体同时经PCRHpDNA 26kDa26kDapET32a( )Bam、HIHindⅢ双酶切、纯化;T4连接酶连接;转染大肠菌以及筛选含插入片段的重组载体。Top10结果:经酶切、测序分析插入的基因 片段为表达外膜蛋白基因,有发生变异,以及氨基酸残基改变。与文献相比:有高度的同源性。Hp26kDa1.1%bp1.51%结论:外膜蛋白的基因克隆、重组载体的构建将有可能为一种有效蛋白质疫苗的制备打下基础。 Hp

The cloning of 26kDa outer membrane protein gene of Helicobacter pylori

Objective: To construct a recombinant vector which could express 26kDa outer membrane protein(OMP) from Helicobacter pylori(Hp) ,and exploit the possibility obtaining the vaccine conferring protection from Hp infection and a diagnostic reagent kit quickly detecting Hp infection. Methods: The gene encoding the structural 26kDa OMP of Hp was amplified from Hp chromosomal DNA by PCR, and inserted in the prokaryotic expression vector pET32a( ), and transformed into the Top10 E. coli strain. Recombinant vector was selected and identified to be transformed into BL-21(DE3) E. coli strain and expressed recombinant protein. Results: The cloned gene of 26kDa OMP was amplified to be 594 base pairs(bp), 1.1% of the cloned gene was mutated , 1.51% of amino acid radicles was changed. Compared with reports: there was high homogenicity between them. Conclusion: This result suggested that recombinant vector expressing 26kDa OMP may be an potential source obtaining the vaccine conferring protection from Hp infection and a diagnostic reagent kit quickly detecting Hp infection.