title: [Cloning of human vascular endothelial growth factor cDNA and its expression in COS-7 cells]
abstract: OBJECTIVE: To clone vascular endothelial growth factor (VEGF) cDNA gene, construct its eukaryotic expression vector and to express this recombinant plasmid in COS-7 cells. METHODS: Human VEGF165 cDNA was amplified by RT PCR from human ovarian carcinoma. After DNA sequenced, the VEGF165 cDNA was inserted into eukaryotic expression vector pcDNA3.1(-). The recombinant plasmid pcDNA3.1 VEGF165 containing VEGF165 cDNA was identified by enzyme digestion and transferred into COS-7 cells mediated by liposome. The transient expression of VEGF was detected by immunohistochemical staining. RESULTS: The cloned VEGF165 cDNA was confirmed by enzyme digestion and DNA sequence analysis. The immunohistochemical results showed that the VEGF165 protein was expression in COS-7 cells 72 h after gene transfer. CONCLUSION: VEGF165 cDNA gene successfully cloned and expressed in COS-7 cells.
authors.affiliation: The Second Affiliated Hospital, College of Medical Sciences, Zhejiang University, Hangzhou 310009, China.
authors.forename: Yi-Xiong
authors.initials: YX
authors.lastname: Zheng
authors.forename: Li
authors.initials: L
authors.lastname: Chen
authors.forename: Shao-Ming
authors.initials: SM
authors.lastname: Xu
authors.forename: Jia-Ping
authors.initials: JP
authors.lastname: Peng
authors.forename: Jiang
authors.initials: J
authors.lastname: Cao