Prokaryotic expression and bioactivity of human platelet-derived growth factor B chain mature peptide.
- Author:
Chun-lu YANG
1
;
Jian-ting CHEN
;
Fan DENG
;
Jian-jun WANG
;
Shen-qiu LUO
;
Da-di JIN
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cell Proliferation; Cells, Cultured; DNA Replication; Escherichia coli; genetics; Genetic Vectors; Humans; Osteoblasts; metabolism; Proto-Oncogene Proteins c-sis; biosynthesis; genetics; Rats; Rats, Sprague-Dawley; Recombinant Proteins; biosynthesis; genetics
- From: Journal of Southern Medical University 2008;28(2):166-168
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo express human platelet-derived growth factor (hPDGF) B chain mature peptide gene in a prokaryotic expression system and detect the bioactivity of the expressed protein.
METHODShPDGF B chain mature peptide gene was amplified and expressed in E. coli, and the recombinant protein, rhPDGF-BB, was purified and renatured in GSSG/GSS system. The bioactivity of rhPDGF-BB in vitro was evaluated with SD rat osteoblasts.
RESULTSThe full-length PDGF-B mature peptide gene was obtained and verified, and successfully expressed in E. coli. Bioactivity detection results showed that the expressed rhPDGF-BB obviously promoted the proliferation and DNA replication of SD rat osteoblasts in vitro (P<0.01).
CONCLUSIONhe PDGF-B chain mature peptide cDNA has been successfully cloned and the PDGF-B precursor highly expressed in E. coli, and renatured rhPDGF-BB displays high bioactivity as shown by MTT assay and flow cytometry. This success provides the basis for production of functional PDGF-BB and facilitates further studies of its role in fracture healing and trauma reconstruction.
