[Proteomic analysis of different fractions in M. tuberculosis culture].
To provide a basis of molecular genetic analysis of M. tubereulosis, the proteomic profiling was prompted. M. tuberculosis H37RV was cultured in Sauton medium at 37 degrees C for 3 weeks, harvested and fractionated into three portions: suspension filtration proteins(A), cytosol proteins(B) and membrane proteins(C). These fractions were analyzed by pH3-10 IPG gradient and SDS-PAGE. The silver-stained technigue and gel images were used. Then the image was transfered into 2-DE gel analysis Software. A part of protein sports of expression level higher from the culture filtrate fraction were identified by peptide mass fingerprinting. A total of 907, 884 and 681 protein sports were observed for A. B. C fractions in M. tuberculosis H37RV, respectively. Distribution of proteins mass for 3 fractions were principally similar, About 70.5-74.4 per cent were distributed in the ranges of Mr 10-49 kD.pI of the proteins for A, B fractions were pricipally similar, About 80.9-83.5 per cent were distributed in the ranges of pH 3.0-6.4, But the number of protein sports for C fraction distributed in the ranges of pH 7.6-10.0 were more than A, B fractions. The number of protein sports of expression level higher for A, B and C fractions were 71(7.8%), 242(27.4%), 19 (2.8%), respectively. 90 pen cent from them, pH of the proteins were distributed in the ranges of pH 3.0-6.4. 73.1 per cent for proteins mass of C fraction were distributed in the ranges of 10 kD-49 kD, which were more than A, B fractions. Nine of the proteins identified ih this study appeared to be homology or putative fanction proteins, but another five proteins were unknown. The proteomic profiling of different fractions in M. tuberculosis obtained in here will be provide a basis for detailed analysis of biology functions of the proteins.
Zhuang
Yuhui
Y
Tuberculosis Research Laboratory, The 309th Hospital of PLA, Beijing 100091, China.
He
Xiuyun
X
Zhang
Xiaogang
X
Li
Guoli
G
Que
Haiping
H
Liu
Shaojun
S
D001426:Bacterial Proteins / Q000032:analysis*; D002462:Cell Membrane / Q000737:chemistry; D003470:Culture Media / Q000737:chemistry; D003593:Cytoplasm / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006863:Hydrogen-Ion Concentration; D009169:Mycobacterium tuberculosis / Q000737:chemistry*; D010449:Peptide Mapping; D016133:Polymerase Chain Reaction; D020543:Proteome / Q000032:analysis*
D001426:Bacterial Proteins; D003470:Culture Media; D020543:Proteome