Construction of the Eukaryotic Expression Plasmid of S677 Mutant MORC2 with Flag Tag and its Expression

The eukaryotic expression plasmids of the pcDNA3.1-MORC2(WT/S677A/S677E) with Flag tag were constructed and explored its expression in SGC-7901 cells,which will facilitate further functional study on MORC2 and its 677 mutant site.First,the pcDNA3.1-Flag vector was acquired with multiple cloning site containing KpnⅠ and XhoⅠ restriction endonuclease.Then,the coding sequence of MORC2 and its mu-tants were acquired from the plasmid pcDNA3.1A-MORC2-WT/S677A / S677E and subcloned into pcD-NA3.1-Flag expression vector by KpnⅠ and XhoⅠ to construct the eukaryotic expression plasmids pcD-NA3.1-Flag-MORC2(WT/S677A/S677E).After the restriction endonuclease digestion and DNA sequencing confirmation,the recombinant plasmids were transfected into SGC-7901 cells and the protein expression were identified by Western blot using flag-tagged antibody.The human MORC2 and its mutants recombinant plasmids with flag tag were obtained successfully and the fusion proteins were expressed successfully in SGC-7901 cell,which provide the basis for the further study on the biology functions of MORC2.