Skip to main content
. 2020 Jul 16;12(7):767. doi: 10.3390/v12070767

Figure 4.

Figure 4

Generation of the E1-deleted CAdV2 mutant. The E1 region of pBAC-CAdV2 was replaced with the Venus expression cassette. The galK and kanamycin resistance gene (galK-Kn) expression cassette, flanked by homologous region 1 (H1) and region 2 (H2), was generated by PCR and introduced into SW102 cells harboring pBAC-CAdV2. After homologous recombination, cells were selected by GalK and Kn, whereupon those harboring pBAC-CAdV2-ΔE1-GalK-Kn were obtained. The Venus expression cassette flanked by H1 and H2 was generated by PCR and introduced into the SW102 cells harboring pBAC-CAdV2-ΔE1-GalK-Kn. After homologous recombination, cells were selected by GalK, whereupon those harboring pBAC-CAdV2-ΔE1-Venus were obtained. CAdV2-ΔE1-Venus was rescued through the transfection of restriction enzyme-digested pBAC-CAdV2-ΔE1-Venus into MDCK-CAdV2-E1-26 cells. After the MDCK cells had been infected with CAdV2-ΔE1-Venus at an MOI of 0.4, Venus expression was observed by fluorescence microscopy. The image was taken at 24 h post-infection.