不同信号肽对重组B群脑膜炎奈瑟菌H因子结合蛋白表达的影响

Effect of various signal peptides on expression of recombinant serogroup B Neisseria meningitidis factor H-binding protein

目的探讨不同信号肽对重组B群脑膜炎奈瑟菌H因子结合蛋白(factor H-binding protein,f HBP)表达的影响。方法运用PCR方法将B群脑膜炎奈瑟菌f HBP原始信号肽分别替换为大肠埃希菌主要外膜脂蛋白(major outer membrane lipoprotein,MLP)、脂蛋白-28(lipoprotein-28,LP)和流感嗜血杆菌(Haemophilus influenzae)P4外膜蛋白信号肽,并将其分别连接至p ET-30a表达载体,转化入大肠埃希菌感受态细胞,IPTG诱导表达,SDS-PAGE鉴定蛋白表达水平。结果替换了P4、LP和MLP信号肽的重组B群脑膜炎奈瑟菌f HBP表达量较带原始信号肽的B群脑膜炎奈瑟菌f HBP相比,分别提高了2.68、1.94和2.91倍。结论替换MLP信号肽对重组B群脑膜炎奈瑟菌f HBP蛋白表达的影响最大,为提高B群脑膜炎奈瑟菌f HBP表达量的研究工作提供了新思路。

Objective To investigate the effect of various signal peptides on expression of factor H-binding protein（f HBP）of serogroup B Neisseria meningitides. Methods The original signal peptide of f HBP of serogroup B N. meningitides was substituted with the major outer membrane lipoprotein（MLP）, lipoprotein-28（LP） and P4 outer membrane protein of Haemophilus influenzae by PCR respectively, and inserted into vector p ET-30 a. The constructed recombinant plasmids were transformed to competent E. coli BL21（DE3）for expression under induction of IPTG. The expression levels of target proteins were determined by SDS-PAGE. Results The expression levels of f HBP of serogroup B N. meningitides in which original signal peptide were substituted with P4, LP and MLP increased by 2. 68, 1. 94 and 2. 91 folds compared with that before substitution, respectively. Conclusion MLP showed significant effect on the expression of f HBP of recombinant N. meningitides, which provided a novel route for study on increasing the expression level of f HBP.