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Data Collection for:
Bartlett et al. (2002). Journal of General Virology, 83, part 8, pp. 1965–1976.
The following material is available only in JGV Online:
 | Fig. 1. (a) Southern blot analysis of virus genomes. VV DNA was digested with HindIII, EcoRI or BglII and fragments were resolved by electrophoresis on an agarose gel (1.0 %, w/v) and transferred to a nylon membrane. Filters were probed with a 925 bp PstI–SstI DNA fragment that contains the N1L ORF, left and right flanking regions and had been labelled by random priming. Bound probe was detected with ECL technology. Size markers are shown in kilobases. (b) Diagram showing the N1L gene locus, including the N1L ORF, relevant restriction sites, the region of DNA deleted in v N1L and the probe used in the Southern blot shown in (a). |
 | Fig. 2. Growth properties of vN1L in vitro. (a) Multiple-step growth curves. BS-C-1 cells were infected at 0.01 p.f.u. per cell. After 1 h, the virus inoculum was removed and the monolayer was washed twice with PBS before the addition of DMEM containing 2.5 % FBS. At various times p.i., the medium was removed and the cells were washed in PBS, scraped from the flask, collected by centrifugation and resuspended in 1 ml 0.1 % BSA in PBS. The samples were frozen and thawed three times, sonicated and the virus infectivity was titrated in duplicate on BS-C-1 cell monolayers. (b) Plaque phenotype. vN1L, vN1l and vN1L-rev were used to infect monolayers of BS-C-1 cells. The infected cells were overlaid with DMEM containing 2.5 % FBS and 1 % carboxymethyl cellulose. After 48 h the culture medium was removed and the cells were stained with 0.1 % crystal violet in 15 % ethanol. |
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