军事医学2017，Vol.41Issue(3)：205-208,245,5.DOI:10.7644/j.issn.1674-9960.2017.03.010
铜绿假单胞菌凝集素PA-ⅠL的纯化表达及活性评价
Expression, purification and biological activity evaluation of Pseudomonas aeruginosa lectin PA-ⅠL
徐茂凯 1孔德聪 1郑玉玲 1郝淮杰 2姜永强1
作者信息
- 1. 军事医学科学院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京 100071
- 2. 中国科学院微生物研究所,病原微生物与免疫学重点实验室,北京 100101
- 折叠
摘要
Abstract
Objective To construct the recombinant plasmid of PA-ⅠL and express in E.coli BL 21 (DE3), and evaluate the biological activity of recombinant protein.Methods PA-ⅠL gene was amplified by PCR using primers designed according to Pseudomonas aeruginosa genome sequences and then cloned to the vector pET -28a ( +).The recombinant plasmid was transformed into E.coli BL21(DE3) and induced to express by IPTG.The recombinant protein was purified by nickel affinity chromatography.The binding activity of recombinant PA-ⅠL with Gb3/CD77 was evaluated by flow cytometry.The function of recombinant PA-ⅠL on the binding of bacteria with host cells was evaluated by colony plate counting.Results and Conclusion The recombinant PA-ⅠL protein was highly expressed in E.coli BL21(DE3) and protein purity by SDS-PAGE analysis was high after nickel affinity chromatography .Besides, the recombinant PA-ⅠL had binding activity to Gb3/CD77 and inhibited the binding of PAO1 to host cells in a dose-dependent manner.关键词
铜绿假单胞菌/PA-ⅠL蛋白/糖鞘脂Key words
Pseudomonas aeruginosa/PA-ⅠL/glycosphingolipid分类
生物科学引用本文复制引用
徐茂凯,孔德聪,郑玉玲,郝淮杰,姜永强..铜绿假单胞菌凝集素PA-ⅠL的纯化表达及活性评价[J].军事医学,2017,41(3):205-208,245,5.
