A recombinant viral clone that fails to express KSHV ORF35 (BAC16-ORF35-stop) was generated.
BAC16-ORF35-stop preserves intact adjacent and overlapping reading frames.
BAC16-ORF35-stop virus displayed significantly reduced lytic virus reactivation.
Exogenous expression of ORF35-Flag reversed the effects of ORF35 deficiency.
ORF35 is important for efficient lytic reactivation of KSHV.