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PMID: 2251260 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

In vitro assembly of infectious nucleocapsids of bacteriophage phi 6: formation of a recombinant double-stranded RNA virus.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 23 ·1990-12-00 ·Pages 9173-7

Olkkonen VM, Gottlieb P, Strassman J, Qiao XY, Bamford DH, Mindich L

Abstract

A system is described for assembling infectious bacteriophage phi 6 nucleocapsids in vitro. Procapsids encoded by cDNA copies of genomic segment L in Escherichia coli were used to package and replicate viral RNA segments. The resulting filled particles were shown to be capable of infecting host cell spheroplasts after incubation with purified nucleocapsid shell protein P8. The infected spheroplasts yielded infectious virions. A modified cDNA-derived RNA segment was inserted into virions by this method. The resulting infectious virions contained the same 4-base-pair deletion as the modified cDNA. These findings support the contention that the preformed procapsids are the "machine" that replicates the phi 6 genome, by showing that the cDNA-derived procapsids are competent to package and replicate RNA properly.

MeSH Terms
Bacteriophages/genetics Base Sequence Capsid/genetics DNA, Viral/genetics DNA-Directed RNA Polymerases/metabolism Molecular Sequence Data Plasmids Polymerase Chain Reaction Pseudomonas/genetics RNA Viruses/genetics RNA, Double-Stranded/genetics RNA, Viral/genetics Restriction Mapping Viral Core Proteins/genetics
Chemicals
DNA, Viral RNA, Double-Stranded RNA, Viral Viral Core Proteins DNA-Directed RNA Polymerases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Olkkonen V M
Department of Genetics, University of Helsinki, Finland.
Gottlieb P
Strassman J
Qiao X Y
Bamford D H
Mindich L
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38 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.87.23.9173
Published
1990-12-00
Pages
9173-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC55126
Subset
IM
Grants
NIGMS NIH HHS · GM31709 · United States
External Links
PubMed source DOI: 10.1073/pnas.87.23.9173 PMC Full Text View journal details
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