[Construction and identification of yeast expression vectors containing human platelet-derived growth factor B-chain gene].
OBJECTIVE
To investigate the expression efficiency of human platelet-derived growth factor B-chain (PDGF-B) gene in yeast and assess the activity of the expressed product.


METHODS
A full-length complementary DNA of human PDGF-B gene was amplified from the total RNA extracted from human vascular endothelial cells using reverse transcription (RT)-PCR and then cloned into pGEM-T vector. The PCR products with specific primers were recombined into yeast expression plasmids pMETB or pMETalphaA, followed by identification with restriction endonuclease.


RESULTS
The 578 bp fragment encoding mature PDGF-BB peptide with signal peptide and the 340 bp fragment without signal peptide were identified and verified by restriction endonuclease mapping and sequencing, and two yeast expression vectors pMETB-PDGFB(1) and pMETalphaA-PDGFB(2) were reconstructed.


CONCLUSIONS
The yeast expression vectors containing human PDGF-B gene have been successfully constructed for further studies of gene expression in yeast.
Yang
Chun-lu
CL
Department of Orthopedic Surgery, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China. ycl_2002@163.com
Chen
Jian-ting
JT
Deng
Fan
F
Tang
Huan-zhang
HZ
Tan
Xiao-yun
XY
Luo
Shen-qiu
SQ
D000077214:Becaplermin; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D010982:Platelet-Derived Growth Factor / Q000235:genetics*; D020574:Proto-Oncogene Proteins c-sis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015003:Yeasts / Q000235:genetics*
D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D000077214:Becaplermin