Construction and expression of eukaryotic expression vector containing hVEGF165 gene in rat bone marrow stroma cell

• Published in: Xi bao yu fen zi mian yi xue za zhi Vol. 19; no. 1; p. 32
• Main Authors: Gao, Quan-wen, Dong, Shao-zhong, Chen, Xi-zhe, Chen, Yan, Yang, Lian-jia
• Format: Journal Article
• Language: Chinese
• Published: China 01.01.2003
• Subjects: Animals; Bone Marrow Cells - metabolism; Escherichia coli - genetics; Escherichia coli - metabolism; Genetic Vectors; Humans; Rats; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Stromal Cells - metabolism; Transfection; Vascular Endothelial Growth Factor A - genetics; Vascular Endothelial Growth Factor A - metabolism
• ISSN: 1007-8738

To construct the eukaryotic expression vector containing human vascular endothelial growth factor 165(VEGF165) gene and express it in rat bone marrow stroma cells(rMSCs). The recombinant plasmid pSP73-VEGF165 was digested with BamH I and Xho I. Then the hVEGF165 gene segment obtained was again cloned into pcDNA3.1 to construct recombinant eukaryotic expression vector pcDNA3.1-VEGF165. Then the recombinant vector was identified by enzyme digestion analysis and sequencing. The rMSCs were transformed by recombinant vector and positive clones were screened with G418. The expression of hVEGF165 gene in the transformed cells was detected by immunocytochemical staining. Enzyme digestion analysis and sequencing showed that target gene had been cloned into recombinant vector. The expression of hVEGF165 gene in the transformed cells had been demonstrated by immunocytochemical staining. The recombinant eukaryotic expression vector has been constructed and expressed successfully in the transformed cells. Therefore, it is