牙龈卟啉单胞菌红血球凝集素B蛋白纯化和鉴定

Purification and idenfication of recombinant HagB of porphyromonas gingivalis

目的:牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)是牙周炎的重要病原菌。该菌膜表面蛋白-红血球凝集素参与细菌黏附和入侵宿主细胞的过程。本研究目的是纯化和鉴定Pg381膜表面红血球凝集素-B(hemagglutinin B,HagB),为能进一步研究该蛋白的特性做准备。方法:HagB基因构建在载体pQE31上,用E.coliM15蛋白表达株通过Ni-NTA亲和柱来表达,表达带有Histag的目的蛋白,做蛋白测序鉴定,制备并筛选相应的抗体。结果:本实验通过分子生物学技术,纯化出目的蛋白HagB,经过抗原免疫,得到相应的单克隆抗体。结论:本实验中的E.coliM15[pREP4]pQE-31-TX1大肠杆菌表达系统能有效地表达HagB蛋白。

AIM： Porphyromonas gingivalis is a periodontopathogen. Hemagglutinins may function as adhesins and invsions, and are required for virulence of several bacterial pathogens. The aim of this study was to purify the protein of hemagglutinin B （HagB） of P. g381. METHODS ： The hagB gene of P. g381 （ 1.4 kb） was cloned into the vector pQE31 and the construct was designated pQE31 -TX1. The histidine -tagged HagB was purified on a nickel - nitrilotriacetic acid （ Ni - NTA） affinity column with Fast Protein Liquid Chromatography from E. coli M15 [ pREP4] pQE -31 -TX1. The purified protein was sequenced and used to produce monoclone antibody of HagB. RESULTS： The recombinant hemagglutinin B protein of P. g 381was expressed in E. coli M15 [ pREP4 ] pQE - 31. Protein sequences confirmed the identity of the purified protein as being hemagglutinin B of P. g381. CONCLUDION： E. coli M15 [ pREP4 ] pQE- 31 -TX1 can provide hige productive recombinantion HagB protein.