Construction and expression of eukarª²yotic expression vector of fusion gene DDR2ª²Fc

    ZHANG Yan, WANG Baoª²Li, SU Jin, CHE Hongª²Lei, YU Jiangª²Tian, LIU Xinª²Ping

    College of Life Sciences, Northwest Sciª²tech University of Agriculture £¦ Forestry, Yangling 712100, China, Department of Biochemistry £¦ Molecular Biology, School of Basic Medicine, Fourth Military Medical University, Xi¬ðan 710033, China

    ¡¾Abstract¡¿ AIM: To construct the eukaryotic expression vector of DDR2ª²Fc fusion gene and to investigate its expression in HEK293 cells. METHODS: The two cDNAs was amplified by PCR respectively from the rat brain tissue and the plasmid conª²taining the fullª²length cDNA of human IgG1 Fc,and cloned to the eukaryotic expression vector pcDNA3.1(-) by directional cloning. The resultant recombinant plamid pcDNA3.1(-)/DDR2ª²Fc was transfected into HEK293 cells with liposome transfection reagent. Then RTª²PCR, Western Blot were used to detect the expression of the fusion protein. RESULTS: DNA sequencing and restriction enzyme digestion verified the correction of recombinant plasmid pcDNA3.1(-)/DDR2ª²Fc. The expressed fusion protein was detected in the transfected HEK293 cells and the molecular weight of the protein was the same as we expected. CONCLUSION: The recombinant plasmid pcDNA3.1(-)/DDR2ª²Fc was successfully constructed and the fusion protein DDR2/ Fc was expressed in HEK293 cells. ......