Drug-resistant gene of mycobacterium tuberculosis and application of drug-resistant gene of mycobacterium tuberculosis

A technology of Mycobacterium tuberculosis and drug resistance, applied in the application, antibacterial drugs, genetic engineering and other directions, can solve problems such as unexplainable drug resistance, and achieve the effect of convenient anti-tuberculosis performance

Active Publication Date: 2015-03-18
SHANGHAI PULMONARY HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although many drug resistance genes against a single anti-tuberculosis drug have been revealed, the known drug resistance genes can only explain part of the drug resistance of clinical strains, and cannot explain all drug resistance, especially the multidrug resistance of clinical strains

Method used

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  • Drug-resistant gene of mycobacterium tuberculosis and application of drug-resistant gene of mycobacterium tuberculosis
  • Drug-resistant gene of mycobacterium tuberculosis and application of drug-resistant gene of mycobacterium tuberculosis
  • Drug-resistant gene of mycobacterium tuberculosis and application of drug-resistant gene of mycobacterium tuberculosis

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Effect test

Embodiment 1

[0030] 1. Materials and methods

[0031] 1. Axgen plasmid extraction kit, TakaRa’s PMD18-T vector kit, Beijing Tiangen’s DNA gel recovery kit, TakaRa’s high-fidelity enzyme,

[0032] 2. Method

[0033] 2.1 Isolation of Mycobacterium tuberculosis containing MFG71 / 74-9 gene and drug sensitivity detection of first-line anti-tuberculosis drugs

[0034] Add an equal volume of 4wt% NaOH solution to the sputum samples from tuberculosis patients in Shanghai Pulmonary Hospital, vortex for 15-20 minutes, then add 20-40mL of 0.067mol / L PBS and mix well. Centrifuge at 3000g for 30min, remove the supernatant, add 0.5mL PBS to the precipitate and mix well; take 0.1ml of the treated specimen, inoculate acidic Roche medium aseptically, culture at 37°C until the colony grows, and then follow the basic professional committee of the Chinese Antituberculosis Association The "Tuberculosis Diagnosis Laboratory Inspection Regulations" edited for strain identification and drug susceptibility testin...

Embodiment 2

[0052] Embodiment 2, bacterial plasmid transduction experiment and drug resistance detection

[0053] 1. Materials

[0054] E.coli DH5α; Mycobacterium smegmatis (ATCC19420); Axgen plasmid mini-prep kit; TakaRa company's PMD18-T vector kit; E.coli-mycobacterium shuttle plasmid PVV16; Beijing Tiangen company's DNA gel Recovery kit, TakaRa's endonuclease BamHI and HindIII TakaRa's T4DNA ligase and high-fidelity enzyme.

[0055] 2. Method

[0056] 2.1 After extracting the PMD18-T-MFG71 / 74-9 recombinant plasmid obtained in Example 1 with a plasmid extraction kit, perform double digestion with BamHI and HindIII with PVV16 at the same time, and use the DNA gel recovery reagent for the digested product The cassette was recovered, and then ligated by T4 DNA ligase to obtain the PVV16-MFG71 / 74-9 recombinant plasmid, which was transformed into E.coli DH5α, and positive clones were screened by culturing on a plate containing Kan resistance at 37°C. The recombinant plasmid PVV16-MFG71 / 7...

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Abstract

The invention provides a drug-resistant gene of mycobacterium tuberculosis and an application of the drug-resistant gene of the mycobacterium tuberculosis. The drug-resistant gene is used for coding any one of the following proteins: i) a protein sequence including an SEQ ID No. 2 sequence in an amino acid and / or nucleotide sequence table; and ii) a protein sequence derived by replacing, deleting or superposing one or more amino acids in sequence (i), and provided with the same function as sequence i). According to a drug sensitivity test, the drug-resistant gene can be used for enabling bacteria to generate drug resistance to various anti-tuberculosis drugs.

Description

technical field [0001] The invention relates to a gene fragment, in particular to a new drug resistance gene of Mycobacterium tuberculosis. Background technique [0002] Tuberculosis is a chronic infectious disease that seriously endangers people's health. Among them, Mycobacterium tuberculosis (M. tuberculosis, commonly known as Mycobacterium tuberculosis) is the pathogenic bacteria that causes tuberculosis. Mycobacterium tuberculosis can invade all organs of the body, but tuberculosis is the most common. my country is one of the 22 countries with severe tuberculosis epidemics in the world. Although, with the improvement of living standards and health conditions, especially the implementation of group prevention and group treatment, children are generally vaccinated against BCG, which greatly reduces the incidence and mortality of tuberculosis. However, the emergence of drug-resistant and multidrug-resistant Mycobacterium tuberculosis has made the situation of tuberculosis...

Claims

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Application Information

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IPC IPC(8): C12N15/31C07K14/35C07K16/12C12N15/74A61K39/04A61P31/06
Inventor 秦莲花戈宝学胡忠义王洁陆俊梅
Owner SHANGHAI PULMONARY HOSPITAL
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