[Construction of eukaryotic expression vector for VEGF165 gene and its expression in rat bladder smooth muscle cells].
OBJECTIVE
To construct an eukaryotic expression vector for vascular endothelial growth factor (VEGF) 165 gene and obtain VEGF expression in rat bladder smooth muscle cells.


METHODS
VEGF165 cDNA was cloned into the eukaryotic expression vector pcDNA3.1(-), and the resultant recombinant vector pcDNA3.1(-)/VEGF165 was transfected into the rat bladder smooth muscle cells by electroporation. VEGF expression in the cells was determined by RT-PCR and immunofluoresence assay, and the biological activity of VEGF in the supernant of the transinfected cell culture was tested by MTT assay.


CONCLUSIONS
VEGF expression was obtained in the transinfected cells, and the supernant of the transinfected cell cultures stimulated the proliferation of the endothelial cells.
Lin
Mao-hu
MH
Postgraduate School, Fourth Military Medical University, Xi'an 710032, China.
Zhao
Shan
S
Miao
Rui
R
Jia
Ning
N
Li
Juan
J
D000818:Animals; D000831:Animals, Newborn; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003001:Cloning, Molecular; D017077:Culture Media, Conditioned / Q000378:metabolism / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics*; D005057:Eukaryotic Cells / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D001743:Urinary Bladder / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology
D017077:Culture Media, Conditioned; D018076:DNA, Complementary; D042461:Vascular Endothelial Growth Factor A