Fig. 4.

Identification of DEV CHv-BAC-GΔUL55 and its revertant DEV CHv-BAC-GΔUL55R. a Identification of DEV CHv-BAC-GΔUL55 and DEV CHv-BAC-GΔUL55R after two rounds of RED recombination. Lane1, 4: The product of UL55 region before RED recombination; Lane 2: The product of UL55 region after 1st round of recombination for constructing UL55 deletion mutant. Lane 3, 5: The product of UL55 region after 2nd round of recombination for constructing UL55 deletion mutant; Lane 6, 7: Identification of 1st and 2nd round of RED recombination for constructing UL55 deletion revertant DEV CHv-BAC-GΔUL55R, respectively. b, c Restriction fragment length polymorphism (RFLP) analysis of rescued recombinant virus DEV CHv-BAC-G, DEV CHv-BAC-GΔUL55, DEV CHv-BAC-GΔUL55R. (b)(c) Indicated the orientation and real Gel analysis of rescued recombinant virus digested by EcoR I, respectively. △: The different band. d Identification of UL55 deletion mutant and revertant by IFA, DEV CHv-BAC-G infected DEF cells were detected as parental virus. Rabbit anti-UL55 IgG were used as primary antibody. A-D: DEV CHv-BAC-G infected cells. E-H: DEV CHv-BAC-GΔUL55 infected cells. I-L: DEV CHv-BAC-GΔUL55R infected cells