Mouse DND1 Lentivirus Vector Construction And Lentivirus Production

Objective In this study, we will make an attempt to put Dnd1 gene coming from 10.5d's embryo cDNA of mouse into lentiviral vector system, which will establish a strong foundation for working out the biological function of the protein expressed by Dndl.Methods The DNA fragment of Dnd1 coding sequence was amplified from 10.5d's embryo cDNA by PCR with designed primer, and then subcloned into pGC-FU vector with In-Fusion technique to generate the lentiviral expression vector, pGC-FU-Dndl. The positive clones were screened by PCR and the correct Dnd1 was confirmed by sequencing. Recombinant lentiviruses were produced in 293T cells following the cotransfection of pGC-FU-Dndl, and packaging plasmids of pHelper 1.0 and pHelper 2.0. Dndl protein expression in 293T cells was detected by Western blot.Results and Conclusion The cDNA fragment of Dndl gene was successfully amplified from mouse embryo cDNA library by PCR. The lentiviral expression Vector Carrying Dndl was successfully constructed and high titer of lentivirus was successfully produced, and it can establish a foundation for our study the molecular function of Dnd1.