Properties of the bovine viral diarrhoea virus replicase in extracts of infected MDBK cells
Warrilow, D. , Lott, W. B., Greive, S. , & Gowans, E. J. (2000) Properties of the bovine viral diarrhoea virus replicase in extracts of infected MDBK cells. Archives of Virology, 145(10), pp. 2163-2171.
An assay for the bovine viral diarrhoea virus (BVDV) replicase was developed using extracts from BVDV-infected cells. The replicase activity was maximal approximately 8 h post-infection as measured by the generation of a genomic length radiolabelled RNA. Using a semi-denaturing gel system, three virus-specific in vitro radiolabelled nascent RNA species were identified. A fast-migrating RNA was demonstrated to be the double-stranded replicative form (RF). A second form was shown to be a partially single-stranded/partially doublestranded RNA, characteristic of the replicative intermediate (RI). A third form, which was often undetectable, migrated between the RF and RI and was probably genomic viral RNA. The optimal replicase activity was dependent on 5–10mM Mg2+ and although it was also active in 1–2mM Mn2+ it was inhibited at higher concentrations. The optimum KCl concentration for labelling of the RI and RF were different, suggestive of at least two distinct replicase activities. These results are supportive of a semi-conservative model of BVDV RNA replication.
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|Item Type:||Journal Article|
|Keywords:||bovine viral diarrhoea virus (BVDV), in vitro radiolabelled nascent RNA|
|Divisions:||Current > Schools > School of Chemistry, Physics & Mechanical Engineering|
Current > Institutes > Institute of Health and Biomedical Innovation
Current > QUT Faculties and Divisions > Science & Engineering Faculty
|Deposited On:||26 Jul 2012 12:48|
|Last Modified:||26 Jul 2012 13:52|
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