[Molecular cloning and expression of human PDGF-B chain mature peptide gene].
OBJECTIVE
To 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.


METHODS
Constructed 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.


RESULTS
PDGF-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.


CONCLUSIONS
The 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.
Chen
Jian-ting
JT
Department of Spine and Orthopaedic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. chenjt99@sina.com
Yang
Chun-lu
CL
Deng
Fan
F
Tan
Xiao-yun
XY
Tang
Huan-zhang
HZ
Zhang
Zhong-min
ZM
Jin
Da-di
DD
D000077214:Becaplermin; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D066298:In Vitro Techniques; D010982:Platelet-Derived Growth Factor / Q000096:biosynthesis* / Q000235:genetics; D020574:Proto-Oncogene Proteins c-sis / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D011994:Recombinant Proteins; D000077214:Becaplermin