Construction and identification of recombinant lentiviral overexprssion vector for mouse CX3CR1 gene

Objective To construct and identify the recombinant lentiviral overexpression vector for mouse CX3CR1 gene.Methods Primers were designed according to the sequence of mouse CX3CR1 gene in GenBank,based on which CX3CR1 gene was amplified by PCR and cloned into vector pUbi-MCS-EGFP.The constructed recombinant lentiviral plasmid was identified by PCR and sequencing,then transfected to 293T cells.Green fluorescence of recombinant lentivirus was observed by microscopy,by which the virus titer was calculated.Results The PCR product of CX3CR1 gene showed a band at length of 1 000 bp on agarose gel electrophoretic profile.Both PCR and sequencing confirmed that recombinant lentivirus plasmid pUbi-MCS-CX3CR1-EGFP was constructed correctly.Green fluorescence was observed in 293T cells 48 h after transfection,and the titer of lentivirus was 2 × 108TU / ml.Conclusion Recombinant lentiviral overexpression vector for CX3CR1 gene was successfully constructed,which laid a foundation of study on lentivirus-mediated transfection of bone marrow-derived mesenchymal stem cells with CX3CR1 gene as well as homing capacity of the gene.