解放军医学院学报2017，Vol.38Issue(12)：1160-1164,5.DOI:10.3969/j.issn.2095-5227.2017.12.018
抗HU蛋白抗体抑制金黄色葡萄球菌生物膜形成实验
Effect of anti-HU polyclonal antibody on formation of Staphylococcus aureus biofilm
摘要
Abstract
Objective To investigate the effect of antibodies against DNA binding protein HU on the biofilm formation of Staphylococcus aureus (S.arueus).Methods The gene HU was amplified by PCR using S.aureus NCTC-8325 genome DNA as template,and the recombinant vector pET-28a-HU was constructed and transformed into Escherichia coli BL21 (DE3).The expression of HU protein was induced by IPTG and purified by Ni2+ affinity chromatography.The polyclonal antibodies against HU protein (anti-HU) were prepared and purified by Protein A affinity resin.The interaction between HU and anti-HU was measured by ELISA and the function of anti-HU was verified by the biofilm formation assay of S.aureus.Results Recombinant protein HU was expressed in soluble form,and a high titer (＞ 1 ∶ 100 000) rabbit anti-HU polyclonal antibody was successfully prepared.The results of crystal violet staining after biofilm formation showed that the absorbance of anti-HU IgG at OD595nm were 0.052 and 1.220 at concentrations of 75 mg/ml and 0.6 mg/ml,respectively.Confocal assay showed that anti-HU polyclonal antibodies significantly inhibited S.aureus biofilm formation in a concentration-dependent manner.Conclusion Our results present that the biofilm formation of S.aureus is significantly blocked by the specific antibodies against HU portien,which provides a novel alternative strategy for treatment of S.aureus infection.关键词
金黄色葡萄球菌/HU蛋白/多克隆抗体/生物膜Key words
Staphylococcus aureus/HU protein/polyclonal antibodies/biofilm分类
医药卫生引用本文复制引用
李静静,刘方杰,高亚萍,刘成华,刘玉,张军,杨光..抗HU蛋白抗体抑制金黄色葡萄球菌生物膜形成实验[J].解放军医学院学报,2017,38(12):1160-1164,5.基金项目
国家自然科学基金资助项目(31370170)Supported by the National Natural Science Foundation of China(31370170) （31370170）
