Construction and expression of A eukaryotic plasmid encoding murine VCAM-1 in vitro

Objective: To construct a eukaryotic plasmid expressing murine vascular cell adhesion molecule-1(VCAM-1) and test whether the plasmid could be expressed in the eukaryotic cell in vitro.Methods: The full-length cDNA of murine VCAM-1 gene was amplified by RT-PCR from the total RNA isolated from the mouse embryo liver.Then the cDNA was inserted into the eukaryotic-expression vector pcDNA3.1(+),the resultant recombinant plasmid was confirmed by restriction endonuclease and sequencing,then designated as pcDNA-mVCAM.The recombinant plasmid pcDNA-mVCAM was transfected into eukaryotic cell CHO by LIPOFECTIN.Western blot was used to certify whether the purpose protein was correctly expressed in CHO cells.Results: About 1 Kb cDNA of murine VCAM-1 was correctly amplified.The recombinant pcDNA-mVCAM was successfully constructed,and it could be correctly expressed in the CHO cells.Conclusion: This recombinant pcDNA-mVCAM plasmid will provide a basis for further study on the unknown function of VCAM.