Effect of ClpE depletion on the bacterial protein expression profiles of Streptococcus pneumoniae.
- Author:
Qun ZHANG
1
;
Nan-lin YIN
;
Xue-mei ZHANG
;
Hong WANG
;
Xin LIU
;
Ya-li CUI
;
Yi-bing YIN
;
Wen-chun XU
Author Information
- Publication Type:Journal Article
- MeSH: Adenosine Triphosphatases; deficiency; genetics; Bacterial Proteins; genetics; Electrophoresis, Gel, Two-Dimensional; Gene Expression Profiling; Heat-Shock Proteins; deficiency; genetics; Polymerase Chain Reaction; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Streptococcus pneumoniae; genetics; growth & development; physiology
- From: Journal of Southern Medical University 2009;29(8):1533-1537
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effect of ClpE on the protein expression profiles of Streptococcus pneumoniae.
METHODSclpE-deficient Streptococcus pneumoniae strain was constructed by long flanking homology-polymerase chain reaction (LFH-PCR) and identified by PCR and sequencing. The total bacterial proteins were analyzed by two-dimensional gel electrophoresis and imaging analysis, and the differentially expressed protein spots were excised by dot-gel digestion with trypsin. Peptide mass fingerprinting (PMF) was obtained by analysis of the fragment length by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The PMF was analyzed using software to identify the proteins.
RESULTSThe number of matched protein spots of the two gels was 61%. By sequence database searching, 4 out of the 17 differential protein spots were identified, namely hypoxanthine-guanine, pyrrolidone-carboxylate peptidase1, formate-tetrahydrofolate ligase, and bifunctional protein pyrR.
CONCLUSIONclpE gene-deficient Streptococcus pneumoniae expresses fewer kinds of proteins at also lower levels than the wild-type bacteria, suggesting that ClpE allows the bacteria to adapt to different host environments by inducing the expression of special proteins.
