Flag results of GFP in fertilized eggs infected by lentiviral vector contained common carp (Cyprinus carpio L.) IGF2b

Abstract:

The vector pLenti6.3IRESEGFP was established using cloned jellyfish GFP gene and connected Bam HI and NheⅠ restrictionsites. Then, the vector was approved by transfecting the DH52 competent cells, restriction enzyme digestion and sequencing. After that, 1 μg/μL plasmid was connected with lentiviral vector. To explore the GFP marked effect flag on lentiviral vector mediated transgenetic fish production, this article observes the expression of GFP using fluorescence microscope after infecting 293 T cells by lentiviral vectors which contained only GFP and contained both GFP and fuctional gene at cell levels; at vivo level, the expression of GFP during the developmented fertilized fingers of Jian carp which were infected by lentiviral vector containing the GFP and IGF2b . The results showed that: the green fluorescence was found in 293T cells infected by both GFP contained vector and GFPIGF2b contained vector. After 48h, the expression of GFP was also observed when lentiviral vector(LentiIGF2bIRESEGFP) was infecting the undivided fertilized eggs from bright field and dark field. So, the GFP can be used successfully to flag the integrated effect through observing the green fluorescence in vivo when the undivided fertilized egg was infected by LentiIGF2bIRESEGFP. These illustrated that this study may lay a solid foundation for developing the transgenetic fish breeding based on the lentiviral vector contained GFP.