Construction and Identification of the Lentivirus Recombinant Vector of Mouse Selenoprotein mSelK shRNA

• 1、School of Life Science, Liaoning University, Shenyang 110036
• 2、School of Pharmacy, Liaoning University, Shenyang 110036

Abstract：Objective: The construction of the recombinant lentivirus vector of mouse selenoprotein mSelK shRNA is important for the researches on the molecular functions of mSelK, especially in macrophage and other immune cells. Methods: The 1.9kbp insertion fragment of the original pLKO.1-TRC cloning vector was excised by digestion with AgeI and EcoRI, and the annealing ShRNA oligos of mSelK were cloned into the AgeI and EcoRI sites to construct pLKO.1-mSelK-ShRNA vector. Then the constructed pLKO.1-mSelK-ShRNA vector, psPAX2 packaging plasmid and pMD2.G enveloping plasmid were co-transfected into HEK-293T cells to construct the mSelK shRNA lentiviral recombinant vector pLKO.1-mSelK-ShRNA. The recombinant lentivirus vector's mRNA knockdown effect of mSelK in monocyte cell strain Raw264.7 was detected by RT-PCR and western blot. It was shown that the expression of mSelK mRNA in RAW264.7 cells is significantly reduced than the control group and this means that the knockdown effect of the lentivirus recombinant vector is rather high. Conclusion: The lentivirus recombinant vector pLKO.1-mSelK-ShRNA, which could knock down mice selenoprotein mSelK, was successfully constructed using pLKO.1 recombinant lentivirus vector system. And the construction of the recombinant vector makes a foundation for the further study of the molecular function of mice selenoprotein mSelK.

Keywords： molecular biology mouse selenoprotein mSelK shRNA lentivirusvector.