Construction of the expression vector pVAX1-mSLC and its expression in COS-7 cells

Objective: To construct the eukaryotic expression plasmid carrying murine SLC full length gene, and its expression in COS-7 cells. Methods: pMD-mSLC was digested by XbaⅠand BamHⅠ, and then the fragment about 410 bp was recovered by agarose electrophoresis method and subcloned to the responding sites of eukaryotic expression vector pVAX1. After confirmation by colony PCR and HinDⅢ enzyme digestion analysis, positive recombinant plasmid was transfected into COS-7 cells in vitro by electroporation method. Specific protein of mSLC in the cuture supernatant and lysate of transfected COS-7 cells was detected by Western blot analysis. Results: Colony PCR and restrictive enzyme digestion analysis proved that recombinant expression vector pVAX1-mSLC had been constructed successfully. The result of Western blotting analysis showed the mSLC expression in the cuture supernatant and lysate of transfected COS-7 cells. Conclusion: Recombinant eukaryotic expressing plasmid pVAX1-mSLC has been constructed successfully and transiently expressed in COS-7 cells,which lays some foundation for the study of SLC gene therapy in murine tumor models.