Construction of a recombination lentiviral vector carrying EZH2 gene coding region and 3'untranslated region fusion protein and its expression in 293T cells

Objective To construct a recombination lentiviral vector carrying human EZH2 gene coding region and 3'UTR fusion protein and its expression in 293T cells.Methods The lentiviral vector was made by the In-Fusion technology.The lentiviral expression vector was obtained after screening followed by sequencing.The lentiviral vector was used to transfect 293T cells and package virus,and the virus titers were determined.Western blotting was used to detect the expression of mCherry-EZH2 in 293T cells.Results The lentiviral vector was successfully constructed,which was confirmed by the assessment of sequencing.After the vector transfected into 293T cells,the lentiviral particles were successfully packaged.Viral titer was 1.59×109 IU/ml.Western blotting was confirmed that the mCherry-EZH2 expressed in 293T cells.Conclusions The lentiviral expression vector was successfully constructed,which expressed the mCherry-EZH2 in 293T cells stablely.