肺炎链球菌假想蛋白SPD0414亚细胞定位及对细菌黏附侵袭的影响  

Subcellular Localization of Streptococcus Pneumonia Hypothetical Proteins SPD0414 and Its Effect on Adhesion and Invasion of S. pn

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作  者:董杰[1] 周爱娥[1] 崔瑾[1] 董姗姗[1] 姜慧[1] 张雪梅[1] 尹一兵[1] 王虹[1] 

机构地区:[1]重庆医科大学检验系临床检验诊断学教育部重点实验室重庆市,400016

出  处:《医学分子生物学杂志》2011年第5期405-410,共6页Journal of Medical Molecular Biology

基  金:国家自然科学基金(No.30970110),重庆市科委自然科学基金(No.CSTC,2009BB5397)

摘  要:目的 为研究肺炎链球菌假想蛋白SPD0414在肺炎链球菌(S.pn)的亚细胞定位,并初步研究其在S.pn黏附和定植中的作用.方法 通过分别在SPD0414的N端和C端融合绿色荧光蛋白(GFP),共聚焦荧光显微镜观察定位情况.用203野生菌和203△spd0414缺陷菌对鼻咽癌细胞CNE和肺腺癌细胞A549细胞的黏附侵袭实验.体内实验中用203和203△spd0414滴鼻感染BALB/c,在6 h和12 h观察细菌在鼻腔和肺中的载量.结果 无论N端或C端融合的GFP都显示绿色荧光,且整个菌体都显示荧光.与203野生菌相比,203△spd0414缺陷菌对CNE和A549的黏附和侵袭能力均显著下降(P<0.05).滴鼻感染BALB/c小鼠,在6 h和12 h,203△spd0414在鼻腔和肺中的细菌载量也明显低于203野生菌(P<0.05).结论 肺炎链球菌假想蛋白SPD0414定位于细菌的胞质.该蛋白在细菌的定植和侵袭中发挥了重要的作用.Objective To study the subcellular localization of streptococcus pneumonia hypo- thetical proteins SPD0414 and its effect on the adhesion and colonization of S. pn. Methods Two vector allowing C-and N-terminal GFP fusions for SPD0414 were used to construct recombinant pro- teins of SPD0414-GFP and GFP-SPD0414 in R6, and the fluorescence was observed under the con- focal microscopy. CNE and A549 were subjected to the adhesion and invasion assays with 203 and 203 A spd0414 stains. In the in vivo experiment, BALB/c mice were infected nasally, and the bacteria load in the nasal cavity and lung was determined after 6 h and 12 h. Results The whole bacteria had obvious fluorescence in mater C-or N-terminal GFP fusions for SPD0414. Besides, the adhe sion and invasion abilities of 203 Aspd0414 to CNE and A549 cells were significantly reduced as compared with that of 203 (P 〈 0.05) . As compared with 203 infection group, the bacteria load in 203 A spd0414 infection group was significantly reduced also in the nasal cavity and lung at 6 h and 12 h ( P 〈 0. 05 ) . Conclusion Streptococcus pneumonia hypothetical proteins SPD0414 locates in cytoplasm, and is associated with the adhesion and invasion of S. pn.

关 键 词:肺炎链球菌 GFP融合蛋白 黏附侵袭 亚细胞定位 

分 类 号:R378.14[医药卫生—病原生物学]

 

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