FIG. 1.

Schematic diagram of KSHV gB ORF replacement with Kan^r gene. The KSHV gB ORF is 845 aa long (2,535 bp). The ∼1.3-kb Kan^r gene flanked by gB sequence (72 bp) was PCR amplified using primer pair A and B and electroporated into DH10B cells harboring the KSHV genome in a BAC plasmid (BAC36 wt-KSHV) with the chloramphenicol resistance gene (Cap^r) and pGET-rec plasmid with the ampicillin resistance gene (Amp^r). Recombinants were selected with chloramphenicol, ampicillin, and kanamycin. Primers E and F amplify the gB ORF flanking the deleted region, and primers C and D amplify a region within the deleted region of the gB ORF.