1.Comparative studies on the proteomics of Rifampin-dependent and-resistant strains of Mycobacterium tuberculosis
Caie YANG ; Mei DONG ; Tieji KUANG ; Xianghong MENG ; Hong LEI ; Minxia SUN
Medical Journal of Chinese People's Liberation Army 2001;0(09):-
Objective To analyze the dielectrophoresis patterns of the proteome of the Rifampin-dependent and-resistant stains of Mycobacterium tuberculosis,to search and identify the differently expressed proteins,and to provide the proteomic basis for researching the mechanism of anti-tuberculosis drug dependence of M.tuberculosis.Methods The whole somatic proteins were extracted from two strains of M.tuberculosis.The first dimensional ampholine electrophoresis was performed on immobilized pH gradient(IPG) rod gels(pH 4-7).Then the proteins on IPG strips were separated using SDS-PAGE.The stained gels were scanned with image scanner and the images were analyzed by Imagemaster 2D software.The differentially expressed proteins were detected by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS).Results Seven hundred and fifty-three spots were detected in Rifampin-dependent strain of M.tuberculosis,while 584 spots were detected in Rifampin-resistant strain,including 404 match spots(the match rate: 61.5%).As to the expression in Rifampin-dependent strain,7 spots significantly up-regulated and 35 spots down-regulated,6 spots were absent in expression,and 5 spots expressed separately,most of the spots were small molecular proteins.Ten spots were selected to run MS analysis.Nine spots were identified as representing 7 proteins.Conclusion The Rifampin-dependent strain of M.tuberculosis is characterized by a rapid and vigorous growth mainly by means of the differential expression of enzymes related to energy metabolism and fatty acid biosynthesis.