[Prokaryotic expression and bioactivity of human platelet-derived growth factor B chain mature peptide].
OBJECTIVE
To 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.


METHODS
hPDGF 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.


RESULTS
The 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).


CONCLUSIONS
he 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.
Yang
Chun-lu
CL
Department of Orthopedic Surgery, Headquarter Hospital of Liaoning Armed Police Force, Shenyang 110034, China. ycl_2002@163.com
Chen
Jian-ting
JT
Deng
Fan
F
Wang
Jian-jun
JJ
Luo
Shen-qiu
SQ
Jin
Da-di
DD
D000818:Animals; D049109:Cell Proliferation; D002478:Cells, Cultured; D004261:DNA Replication; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D010006:Osteoblasts / Q000378:metabolism; D020574:Proto-Oncogene Proteins c-sis / Q000096:biosynthesis* / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D020574:Proto-Oncogene Proteins c-sis; D011994:Recombinant Proteins