Construction and identification of lentiviral vectors expressing cat pluripotency genes

• 1、College of Animal Sciences, JiLin University, Changchun 130062

Abstract：Objiective This paper aims to explore the construction and packing of lentivirus vectors including cat pluripotency genes Oct4、Sox2、Klf4 and c-Myc. Methods The target genes were obtained from the plasmid containing cat Oct4, Sox2, Klf4 and c-Myc, and were directionally connected to the lentiviral vector LV5 carrying green fluorescence protein (GFP) to construct the reconbinant lentiviral vectors LV-Oct4、LV-Sox2、LV-Klf4 and LV-c-Myc. The production was respectively transformed into competent bacteria. The gene sequences were analyzed after identified by positive clones screening and restrictive enzyme digestion. Lipofectamine2000 was used to transfect 293T cells for packing lentivirus and testing the titer of lentivirus. Results The double enzyme digestion and DNA sequencing analysis revealed that the lentivirus vectors were successfully constructed and packed including Oct4, Sox2, Klf4 and c-Myc. The sequence of gene was consistent with objective sequence and the titer of lentivirus reached over 107TU/ml. Conclusion we have successfully constructed the lentiviral vectors expressing cat pluripotency genes and obtained the packaging cell lines stably producing lentiviral particles.

Keywords： zoology cat lentiviral vector pluripotent gene packing