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Construction and analysis of ORF6-null KSHV genome.

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posted on 2014-06-09, 03:38 authored by Can Peng, Jungang Chen, Wei Tang, Chunlan Liu, Xulin Chen

(A) Schematic diagrams of the structures of ORF6 and its neighboring ORFs in the wild-type and mutant BACs. (B) Confirmation of KSHV ORF6 replacement with Kan/SacB cassette by PCR amplification. The BAC36 (lane 1 and 3) and BACΔ6 (lane 2 and 4) were amplified with flanking PCR primers A and B and internal PCR primers C and D. (C) Confirmation of BAC36 and BACΔ6 genome with BglII digestion and Southern blot analysis. BAC36 (lane 1) and BACΔ6 (lane 2) DNAs were digested with BglII. The digested DNAs were electrophoresed, blotted, and hybridized with Dig-labeled ORF6 or Kan/SacB DNA probes, respectively.

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