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The Function Research of Mycobacterium Tuberculosis with Rv1272 and Rv1273

Author: ChengXuHong
Tutor: ZhaoYingWei
School: Suzhou University
Course: Pathogen Biology
Keywords: Mycobacterium tuberculosis Rv1272 Rv1273 membrane protein MIC EB accumulation growth inhibition ATP hydrolysis
CLC: R378.911
Type: Master's thesis
Year: 2011
Downloads: 30
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Abstract


Objective Study on the relationship of two proteins coded by Mycobacterium tuberculosis Rv1272 & Rv1273 with drug efflux and research other aspects of the function of them.Methods Their structures were analyzed by bioinformatics. The two genes were PCR amplified and cloned into expression vector pMF406 to generate the recombinant plasmids followed by sequence confirmation. The verified recombinants were transformed into M. Smegmatis mc~2155. After investigation the subcellular localization of these two proteins by Western Blotting, MICs (minimal inhibitory concentration) of nine drugs that commonly used were tested by test tube method. On the other hand, the accumulation of ethidium bromide by mycobacteria was measured to indentify whether the two protein Outward transport EB. Its entry was measured at room temperature in top-reading mode at an excitation wavelength of 530 nm and an emission wavelength of 590 nm using a Synergy HT reader. And then we construct the mutation strains to further validation the function of the protein. The next, the susceptibility of the recombinant strains that containing the two genes respectively to ethidium bromide was tested in order to further verify its function.Results The recombinants were successfully constructed. The subcellular localization results identified Rv1272 and Rv1273 as membrane proteins. MICs results showed that compared with the control, the recombinant strains that containing the two genes respectively did not change the resistance of the testing drugs. The accumulation of ethidium bromide by wild type strains of Rv1272 had no significant change with the control. There were significant differences between the wild type strains of Rv1273 and the control, and its mutation no differences with the control. Compared with the control, both in Escherichia coli and M. Smegmatis mc2155, growth of the recombinant strains containing the two genes respectively are inhibited.Conclusion Rv1272 and Rv1273 are membrane proteins. Their relationships with the drug tolerance of Mycobacterium tuberculosis need to be further investigated. Rv1272 may have not matter transport function, and Rv1273 may be an inward material transfer pump, and the energy of transport dependence on the ATP hydrolysis.

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CLC: > Medicine, health > Basic Medical > Medical Microbiology ( pathogenic bacteriology,pathogenic microbiology ) > Pathogenic bacteria > Mycobacterium ( acid-fast bacilli ) > Mycobacterium tuberculosis
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