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【目的】Rv3194c基因编码的是结核分枝杆菌的PDZ信号蛋白,本研究对其是否具有黏附素特性进行了探索。【方法】对Rv3194c进行原核表达,然后Rv3194c蛋白分别与透明质酸、硫酸软骨素、Ι型胶原蛋白在不同温度(37、38、39、40°C)的缓冲液中孵育过夜,然后用Western blot和ELISA分析其上清液中组分含量的变化。【结果】Western blot鉴定结果表明,His-Rv3194c蛋白以可溶形式表达,蛋白质的分子质量大小为35 k Da。Western blot表明,39°C实验组的上清中His-Rv3194c蛋白含量(***P<0.001)显著小于其它实验组(37、38、40°C);ELISA表明,39°C实验组的上清中透明质酸(HA)、硫酸软骨素(CS)、Ι型胶原蛋白(CollagenΙ)的含量(***P<0.001)极显著小于其它实验组(37、38、40°C)。【结论】首次证实了Rv3194c蛋白具有黏附素特性,可成为研发新型抗结核药物的靶点蛋白。
【Objective】 The Rv3194c gene encodes the PDZ signaling protein of Mycobacterium tuberculosis. In this study, it was explored whether or not it has adhesin. 【Method】 Prokaryotic expression of Rv3194c was carried out. Then Rv3194c protein was incubated with hyaluronic acid, chondroitin sulfate and collagen type I in buffer solution at different temperature (37, 38, 39, 40 ° C) blot and ELISA analysis of the supernatant changes in the content of components. 【Result】 The results of Western blot showed that His-Rv3194c protein was expressed in soluble form and the molecular mass of the protein was 35 kDa. Western blot showed that the content of His-Rv3194c in the supernatant of the experimental group at 39 ° C (*** P <0.001) was significantly lower than that of the other experimental groups (37, 38 and 40 ° C) The contents of HA, CS and Collagen I in the supernatant (*** P <0.001) were significantly lower than those in the other experimental groups (37, 38 and 40 ° C). 【Conclusion】 For the first time, Rv3194c protein has adhesin characteristics and can be used as a target protein in the development of new anti-TB drugs.