Molecular cloning and expression of human PDGF-B chain mature peptide gene.
- Author:
Jian-ting CHEN
1
;
Chun-lu YANG
;
Fan DENG
;
Xiao-yun TAN
;
Huan-zhang TANG
;
Zhong-min ZHANG
;
Da-di JIN
Author Information
- Publication Type:Journal Article
- MeSH: Cloning, Molecular; Escherichia coli; genetics; metabolism; Genetic Vectors; Humans; In Vitro Techniques; Platelet-Derived Growth Factor; biosynthesis; genetics; Proto-Oncogene Proteins c-sis; genetics; Recombinant Proteins; biosynthesis; genetics; Transfection
- From: Chinese Journal of Surgery 2004;42(19):1170-1173
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo acquire sufficient PDGF-BB protein and provide the basis for the further studies of its role on the fracture healing and trauma reconstruction and its clinical applications.
METHODSConstructed the prokaryotic expression vector pQE-PDGF-B with the gene rearrangement technique, and the monomeric form of recombinant PDGF-B expressed in E. coli M15.
RESULTSPDGF-B mature peptide gene was inserted into prokaryotic expression vector pQE30, which was confirmed by PCR, enzyme digestion and sequencing identification; the expressed products of pQE-PDGF-B in E. coli showed a single protein on SDS-PAGE, and their expression level was about 15% of the total bacterial protein. The molecular weight of the purified PDGF-B protein was about 15 KDs on SDS-PAGE.
CONCLUSIONSThe construction of recombinant plasmid and preparation of the monomeric protein of PDGF-B provides a solid foundation for further studying the function of PDGF-BB and producing biologically PDGF-BB protein.