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. 2024 Aug 26;20(8):e1012081. doi: 10.1371/journal.ppat.1012081

Fig 3. Construction of ORF48 deletion mutants using the BAC16 system.

Fig 3

(A) Agarose gel electrophoresis of wild-type BAC16 and mutant BAC16-ORF48del. The loading sequence from left to right is 1kb plus DNA ladder, WT BAC16, midrange DNA ladder, BAC16-ORF48del candidates #1, #4 and #6. DNA was digested with NheI for two hours and resolved on a 0.4% agarose gel stained with ethidium bromide. DNA ladder sizes covering the KSHV genome are indicated to the left of the gels. (B) Schematic illustration of the strategy used to generate ORF48 deletion mutants. The ORF48 coding region does not overlap with adjacent ORF47 and ORF49. (C) Analysis of BAC16-ORF48del VS WT BAC16. Deletion of the ORF48 coding region (1.2kb) results in a decreased size of a band from 25,693bp to 24,484bp, as indicated by red arrows. (D) Bright-field view and GFP expression in the established WT, ORF48del#1 and #4.