Structural characterization of RB1BvIL-8ΔsmGFP. (A)
Schematic diagram of the MDV genome showing the region expressing the
vIL-8 gene, the restriction sites used in the deletion of vIL-8, the
smGFP expression cassette used in mutagenesis, and the location of PCR
primers used in the structural characterization of the parental and
mutant viruses. (B) An agarose gel showing the results of PCR
amplification using primers 1 and 4 (shown in panel A) (upper left
panel) and Southern blot hybridization analysis of these products
using the BamHI-L fragment of the MDV genome
(upper right panel), the vIL-8 cDNA (lower left panel), and the smGFP
gene (lower right panel) as probes. The lanes are λd3,
HindIII-digested lambda DNA molecular weight markers;
123, 123-bp DNA ladder markers (GIBCO-BRL); BamL, pUC19 containing the
BamHI-L fragment of the MDV genome (20);
pYH, transfer vector pYH6a; CEF, uninfected CEF DNA; RB, RB1B parental
MDV-infected CEF DNA; V1, RB1BvIL-8ΔsmGFP clone 2-infected CEF DNA;
V2, RB1BvIL-8ΔsmGFP reisolated from spleen cells of infected
chickens; and UA20, DNA from cell line MDCC-UA20, established from an
RB1BvIL-8ΔsmGFP-induced ovarian lymphoma.