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. 2009 Aug 5;83(20):10582–10595. doi: 10.1128/JVI.00497-09

FIG. 2.

FIG. 2.

Construction and analysis of ORF33-null MHV-68 (BAC) DNAs. (A) Nucleotide sequence of the region containing the mutation in the ORF33-null MHV-68 mutant. The genome coordinates are given to the left of the nucleotide sequence and on top of the nucleotides between which mutations were introduced. The ORF33 ATG and the three inserted nonsense and frameshift mutations are boxed, and the introduced VspI site is underlined. (B) Schematic representation of the ORF33 locus and its flanking ORFs on the WT BAC or 33STOP genome, the position of a probe (nt 49109 to 50264), and the predicted restriction fragments detected by the probe in Southern blotting analysis. (C) Southern blotting analysis of WT BAC and 33STOP DNAs. Various BAC DNAs were digested with VspI or BglII as indicated at the top of the panel, electrophoresed, blotted, and hybridized with the DIG-labeled probe. Positions of a λ/HindIII DNA ladder run on the same gel are indicated to the left of the panel.