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热休克蛋白与寻常性炎性痤疮:痤疮丙酸杆菌GroEL和DnaK同源物的分子克隆、过表达及纯化

Heat shock proteins and inflammatory acne vulgaris: molecular cloning, overexpression and purification of a propionibacterium acnes GroEL and DnaK homologue.

作者信息

Farrar M D, Ingham E, Holland K T

机构信息

Skin Research Centre, Division of Microbiology, School of Biochemistry and Molecular Biology, University of Leeds, LS2 9JT, Leeds, UK.

出版信息

FEMS Microbiol Lett. 2000 Oct 15;191(2):183-6. doi: 10.1111/j.1574-6968.2000.tb09337.x.

Abstract

Propionibacterium acnes is associated with inflammatory acne. The genes encoding two putative mediators of inflammation, the heat shock proteins GroEL and DnaK, were cloned from this organism and sequenced. groEL and dnaK encode proteins of 56.8 and 66.4 kDa, respectively, which show a high degree of homology (>75% similarity) to the GroEL and DnaK proteins of mycobacteria and streptomycetes. The promoter regions of both genes contain inverted repeat sequences believed to be involved in the transcriptional regulation of heat shock genes. Recombinant P. acnes GroEL and DnaK were overexpressed in Escherichia coli with C-terminal histidine tags. The recombinant proteins were purified from E. coli by metal affinity chromatography. These proteins will now be used in immunological investigations to determine their role in inflammatory acne.

摘要

痤疮丙酸杆菌与炎性痤疮相关。从该生物体中克隆并测序了编码两种假定炎症介质即热休克蛋白GroEL和DnaK的基因。groEL和dnaK分别编码56.8 kDa和66.4 kDa的蛋白质,它们与分枝杆菌和链霉菌的GroEL和DnaK蛋白具有高度同源性(>75%相似性)。两个基因的启动子区域都含有反向重复序列,据信这些序列参与热休克基因的转录调控。重组痤疮丙酸杆菌GroEL和DnaK在带有C端组氨酸标签的大肠杆菌中过表达。通过金属亲和层析从大肠杆菌中纯化重组蛋白。这些蛋白质现在将用于免疫学研究,以确定它们在炎性痤疮中的作用。

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