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PMID: 2989815 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Expression of a functional influenza viral cap-recognizing protein by using a bovine papilloma virus vector.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 82 ·No. 13 ·1985-07-00 ·Pages 4326-30

Braam-Markson J, Jaudon C, Krug RM

Abstract

The gene for the influenza viral PB2 protein, which recognizes and binds the 5'-terminal cap 1 structures (m7GpppNm) on eukaryotic mRNAs, was inserted into a bovine papilloma virus vector under the control of a mouse metallothionein I (MT-I) promoter. After transfection of this vector into mouse NIH 3T3 cells, a cell line, cPB2, was obtained that produces PB2-specific mRNA and authentic PB2 protein. Induction of the MT-I promoter with CdCl2 causes about a 10-fold increase in PB2 mRNA and protein levels. The expressed PB2 protein is functional, as it relieves the block in viral mRNA synthesis exhibited by a temperature-sensitive viral mutant containing a cap-binding defect in the PB2 protein. This demonstrates complementation of a function of a negative-strand RNA virus by a gene product expressed in a cell line from recombinant DNA.

MeSH Terms
Animals Bacterial Proteins Bovine papillomavirus 1/genetics Cell Line DNA Restriction Enzymes/metabolism Deoxyribonuclease HindIII Deoxyribonucleases, Type II Site-Specific Electrophoresis, Polyacrylamide Gel Gene Expression Regulation Influenza A virus/genetics Mice Papillomaviridae/genetics RNA Caps/metabolism Transfection Viral Proteins/genetics,metabolism
Chemicals
Bacterial Proteins RNA Caps Viral Proteins DNA Restriction Enzymes Deoxyribonuclease HindIII BglII endonuclease Deoxyribonucleases, Type II Site-Specific
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Braam-Markson J
Jaudon C
Krug R M
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47 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.82.13.4326
Published
1985-07-00
Pages
4326-30
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC390406
Subset
IM
Grants
NIAID NIH HHS · AI 11772 · United States
NCI NIH HHS · CA 08748 · United States
External Links
PubMed source DOI: 10.1073/pnas.82.13.4326 PMC Full Text View journal details
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