PMID- 15897136
OWN - NLM
STAT- MEDLINE
DCOM- 20070501
LR  - 20161124
IS  - 1000-2588 (Print)
IS  - 1000-2588 (Linking)
VI  - 25
IP  - 5
DP  - 2005 May
TI  - [Prokaryotic expression vector construction, expression and polyclonal antibody 
      preparation of the fusion protein of glutathione S-transferase and peroxisome 
      proliferator-activated receptor-gamma coactivator-1].
PG  - 558-61
AB  - OBJECTIVE: To express the fusion protein of glutathione S-transferase (GST) and 
      peroxisome proliferator-activated receptor-gamma coactivator-1 (PPARgammaC1) in 
      E. coli. and prepare the polyclonal antibody against PPARgammaC1. METHODS: The 
      coding sequence of PPARgammaC1 gene was amplified by reverse transcriptase-PCR 
      (RT-PCR) from the total RNA of Hep G2 cells and inserted into pGEX-4T-1 vector. 
      The recombinant vector was identified by restriction endonuclease digestion 
      analysis and the fusion protein GST-PPARgammaC1 was expressed in E. coli. via 
      IPTG induction. The expressed fusion protein was purified by glutathione-agarose 
      affinity chromatography and used to immunize the egg-laying hens for preparing 
      the polyclonal antibody against GST-PPARgammaC1. RESULTS: Restriction 
      endonuclease digestion analysis demonstrated that the PPARgammaC1 gene had been 
      correctly inserted into pGEX-4T-1 vector, and the expressed fusion protein had a 
      relative molecular mass of approximately 39,000 as shown by SDS-PAGE. The 
      polyclonal antibody obtained from the egg yolk immunoglobulins was found to 
      specifically bind to purified PPARgammaC1 in Western blot analysis. CONCLUSION: 
      The successfully prepared polyclonal antibody against PPARgammaC1 peptide 
      provides a useful reagent for PPARgammaC1 detection.
FAU - Zhang, Yan
AU  - Zhang Y
AD  - Department of Endocrinology, Nanfang Hospital, Southern Medical University, 
      Guangzhou 510515, China. xinchristina@163.com
FAU - Xue, Yao-ming
AU  - Xue YM
FAU - Guo, Ai-lin
AU  - Guo AL
FAU - Zhang, Wen-min
AU  - Zhang WM
LA  - chi
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - China
TA  - Di Yi Jun Yi Da Xue Xue Bao
JT  - Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA
JID - 9426110
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Heat-Shock Proteins)
RN  - 0 (PPARGC1A protein, human)
RN  - 0 (Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Transcription Factors)
RN  - EC 2.5.1.18 (Glutathione Transferase)
SB  - IM
MH  - Antibodies, Monoclonal/*biosynthesis
MH  - Escherichia coli/genetics/metabolism
MH  - Genetic Vectors
MH  - Glutathione Transferase/*biosynthesis/genetics/immunology
MH  - Heat-Shock Proteins/*biosynthesis/genetics/immunology
MH  - Humans
MH  - Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha
MH  - Prokaryotic Cells/metabolism
MH  - Recombinant Fusion Proteins/*biosynthesis/genetics/immunology
MH  - Transcription Factors/*biosynthesis/genetics/immunology
EDAT- 2005/05/18 09:00
MHDA- 2007/05/02 09:00
CRDT- 2005/05/18 09:00
PHST- 2005/05/18 09:00 [pubmed]
PHST- 2007/05/02 09:00 [medline]
PHST- 2005/05/18 09:00 [entrez]
PST - ppublish
SO  - Di Yi Jun Yi Da Xue Xue Bao. 2005 May;25(5):558-61.