微生物學(xué)通報 FEB 20,2010,37(2):312～318

以蛋白激酶G為靶點的抗結(jié)核藥物篩選模型的建立和初步應(yīng)用

邵天舒  魏玉珍  李秋萍  趙莉莉  岑山  余利巖*

(中國醫(yī)學(xué)科學(xué)院 醫(yī)藥生物技術(shù)研究所  北京  100050)

摘  要: 結(jié)核分枝桿菌可以產(chǎn)生11種絲氨酸/蘇氨酸蛋白激酶, 其中蛋白激酶G(PknG)對于結(jié)核分枝桿菌在巨噬細(xì)胞內(nèi)以"持留"狀態(tài)長期存活有著重要作用,。本研究以結(jié)核分枝桿菌基因組DNA為模板, 在大腸桿菌中克隆表達(dá)了MTB PknG蛋白, 并分離純化得到PknG純酶,。本研究還采用三步級聯(lián)反應(yīng)方法測定了PknG酶活性, 建立和優(yōu)化了PknG抑制劑高通量篩選模型,。利用此模型共篩選發(fā)酵液樣品2120個, 化合物樣品2300個, 篩選得到陽性化合物1個, 陽性發(fā)酵液13個, 陽性率0.32%,。

關(guān)鍵詞: 藥物篩選模型, 持留, PknG

The Establishment and Application of Anti-tuberculosis Drugs Screening Model Targeting to PknG

SHAO Tian-Shu  WEI Yu-Zhen  LI Qiu-Ping  ZHAO Li-Li  CEN Shan  YU Li-Yan*

(Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking union Medical College,

Beijing 100050, China)

Abstract: Mycobacterium tuberculosis (MTB) PknG, one of 11 Ser/Thr protein kinases produced by MTB, plays an essential role in the residing of MTB in the macrophages as persistent state. In this work, we ampli-fied the pknG gene from the genome of H37Rv, and constructed a recombinant plasmid pET-pknG to express the PknG protein in the E. coli. Using purified PknG, we established and optimized a high-throughout screening model to screen for PknG inhibitors. Among 2120 samples of microbial fermentations and 2300 samples of compounds tested, one positive compound and 13 positive fermentation samples were identified with inhibitory effect on activity of PKnG, resulting in a 0.32% of positive rate.

Keywords: Drugs screening model, Persistence, PknG

生物谷推薦全文下載：以蛋白激酶G為靶點的抗結(jié)核藥物篩選模型的建立和初步應(yīng)用

更多全文請查看鏈接：http://journals.im.ac.cn

聲明：本文由《微生物學(xué)報》授權(quán)生物谷 www.bioon.com 網(wǎng)站發(fā)布，如需轉(zhuǎn)載請直接與中國科學(xué)院微生物研究所期刊聯(lián)合編輯部聯(lián)系并支付相應(yīng)費用,，未經(jīng)授權(quán)不得轉(zhuǎn)載,，若轉(zhuǎn)載將付相應(yīng)的法律責(zé)任。