Antitubercular drug screening model using tubercle bacillus isocitrate lyase as target

A technology of isocitrate and lyase, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc.

Inactive Publication Date: 2007-08-01
MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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Problems solved by technology

[0010] The present inventor screened new anti-tuberculosis drugs from microbial metabolites by establishing a high-throughput drug screening model targeting Mycobacterium tuberculosis ICL, which has not been reported at home and abroad

Method used

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  • Antitubercular drug screening model using tubercle bacillus isocitrate lyase as target
  • Antitubercular drug screening model using tubercle bacillus isocitrate lyase as target

Examples

Experimental program
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Embodiment 1

[0018] Using the screening model to screen samples of microbial origin and determination method

[0019] 1. Sample source: 200 fermentation broth samples are provided by the strain group of our laboratory every week.

[0020] 2. Sample setting: screen 80 samples per 96-well plate, 80 μl reaction system and 5 μl fermentation broth sample per well; at the same time, set 4 groups of positive controls and 4 groups of negative controls.

[0021] 3. Add 5 μl of the fermentation broth to be screened to the sample wells, add 5 μl of itaconic acid (2 mM) to the positive control wells, and add MOPS buffer (50 mM, pH 7.0) to the negative control wells.

[0022] 4. Add 40 μl of isocitrate lyase solution (enzyme concentration is about 2 μg / ml) to each well with an 8-channel microsampler, shake gently, and mix well.

[0023] 5. Add 40 μl of substrate solution (isocitric acid, MgCl 2 and phenylhydrazine concentrations are 8mM), shake gently, and mix well.

[0024] 6. Place the 96-well pla...

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Abstract

The invention relates to a moudle for sifting antitubercular drugs taking bacillus tubercle citrate lyase as target, and the application of said mould for new antitubercular drugs sifting.

Description

field of invention [0001] The invention relates to an anti-tuberculosis drug screening model targeting isocitrate lyase of Mycobacterium tuberculosis and a method for screening novel anti-tuberculosis drugs targeting persistent bacteria using the screening model. Background of the invention [0002] In recent years, tuberculosis (TB) has made a comeback. About 2 billion people in the world have been infected with Mycobacterium tuberculosis, with 8 million new cases and 2 million deaths every year; nearly 400 million people in my country have been infected with Mycobacterium tuberculosis, and 10% of those infected In the next 10 years, there may be 30 million people with TB, and the control of the tuberculosis epidemic has become a top priority. [0003] Mycobacterium tuberculosis persistence (persistence) and drug resistance (resistance) are two major obstacles to the effective control of tuberculosis. Due to the existence of persistent tuberculosis, the course of tuberculos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/25C12Q1/18
Inventor 肖春玲王旸李大为郝雪秦赵丽洋龚立康张章姚天爵
Owner MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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