[结核分枝杆菌Rv3194c蛋白的表达、纯化和鉴定]。

微生物学报 Pub Date : 2016-12-04

Dongyue Zhao, Lili Lin, Fuli Wen

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引用次数: 0

摘要

目的:利用结核分枝杆菌Rv3194c基因编码PDZ[突触后密度-95 (PSD-95)，果蝇肿瘤抑制蛋白diskslarge-1 (DLG)，紧密连接蛋白occludentes 1 (ZO-1)]信号蛋白，并研究其对结核分枝杆菌的粘附能力。方法:在原核系统中表达Rv3194c蛋白。Rv3194c蛋白分别与透明质酸、硫酸软骨素和胶原蛋白Ι在不同温度(37、38、39、40℃)下孵育过夜。然后用Western blot和ELISA检测培养上清的成分变化。结果:Western blot结果显示，Rv3194c蛋白在原核系统中表达，分子量约为35 kDa，主要以可溶性形式表达。Western blot结果显示，39℃实验组上清中His-Rv3194c蛋白含量明显低于其他实验组(37、38、40℃)(*** p)。结论:本研究首次肯定了检测到黏附活性的Rv3194c蛋白将用于新型抗m抗体的开发。结核病药物。

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[Expression, purification and characterization of Rv3194c protein from Mycobacterium tuberculosis].

Objective: PDZ[Post-synaptic density-95 (PSD-95), Drosophilia tumor suppressor protein diskslarge-1 (DLG), the tight junction protein zonula occludentes 1 (ZO-1)] signal protein was encoded by Rv3194c gene from Mycobacterium tuberculosis, and its ability to adhere M. tuberculosis was studied.

Methods: Rv3194c protein was expressed in prokaryotic system. Rv3194c protein was separately incubated with hyaluronic acid, chondroitin sulfate and collagen Ι overnight at different temperature (37, 38, 39, 40℃). Then component changes of culture supernatant were tested by Western blot and ELISA.

Results: Western blot showed that Rv3194c protein expressed in prokaryotic system, with a molecular weight of about 35 kDa, was mainly in soluble form. Western blot showed that His-Rv3194c protein in supernatant of 39℃ experimental group was significantly less than that of other experimental groups (37, 38, 40℃)(***P<0.001). ELISA showed that hyaluronic acid, chondroitin sulfate and collagen Ι in supernatant of 39℃ experimental group was significantly less than that of other experimental groups (37, 38, 40℃)(***P<0.001).

Conclusion: For the first time it was affirmed that Rv3194c protein with detected activity of adhesions in this study will be targeted to the development of the new anti-M. tuberculosis drug.

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来源期刊

微生物学报

自引率

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发文量

7960

期刊介绍： Acta Microbiologica Sinica(AMS) is a peer-reviewed monthly (one volume per year)international journal,founded in 1953.It covers a wide range of topics in the areas of general and applied microbiology.The journal publishes original papers,reviews in microbiological science,and short communications describing unusual observations. Acta Microbiologica Sinica has been indexed in Index Copernicus (IC),Chemical Abstract (CA),Excerpt Medica Database (EMBASE),AJ of Viniti (Russia),Biological Abstracts (BA),Chinese Science Citation Database (CSCD),China National Knowledge Infrastructure(CNKI),Institute of Scientific and Technical Information of China(ISTIC),Chinese Journal Citation Report(CJCR),Chinese Biological Abstracts,Chinese Pharmaceutical Abstracts,Chinese Medical Abstracts and Chinese Science Abstracts.