caa a22 4500 475766687 CHVBK 20180406123607.0 cr unu---uuuuu 170329e20001001xx s 000 0 eng 10.1007/s007050070046 doi (NATIONALLICENCE)springer-10.1007/s007050070046 Properties of the bovine viral diarrhoea virus replicase in extracts of infected MDBK cells [Elektronische Daten] [D. Warrilow, W. B. Lott, S. Greive, E. J. Gowans] Summary.:   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 double-stranded 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 5-10 mM Mg2+ and although it was also active in 1-2 mM 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. Springer-Verlag/Wien, 2000 Warrilow D. Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Herston, Brisbane, Australia, AU aut Lott W. B. Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Herston, Brisbane, Australia, AU aut Greive S. Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Herston, Brisbane, Australia, AU aut Gowans E. J. Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Herston, Brisbane, Australia, AU aut https://doi.org/10.1007/s007050070046 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s007050070046 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Warrilow D. Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Herston, Brisbane, Australia, AU aut NATIONALLICENCE 700 1- Lott W. B. Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Herston, Brisbane, Australia, AU aut NATIONALLICENCE 700 1- Greive S. Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Herston, Brisbane, Australia, AU aut NATIONALLICENCE 700 1- Gowans E. J. Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Herston, Brisbane, Australia, AU aut Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer