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PMID: 2473440 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Infectious TYMV RNA from cloned cDNA: effects in vitro and in vivo of point substitutions in the initiation codons of two extensively overlapping ORFs.

Nucleic acids research ·Vol. 17 ·No. 12 ·1989-06-26 ·Pages 4675-87

Weiland JJ, Dreher TW

Abstract

Full-length cDNA of the 6.3 kb turnip yellow mosaic virus (TYMV) genome was placed between a T7 promoter and a unique Hind III site. In vitro transcription of Hind III-linearized DNA of clone pTYMC yielded full-length RNA transcripts. In inoculations of Chinese cabbage protoplasts and plants, capped transcripts and virion RNA had similar specific infectivities and produced similar systemic symptoms. We have used the pTYMC clone in studies of the expression of two overlapping open reading frames (1.9 kb and 5.5 kb ORFs) by making mutants with alterations in the initiation codons. Evidence is presented from in vitro translations of mutant and wild type RNAs that both ORFs are expressed from TYMV RNA. A mutant in the initiation codon of the 5.5 kb ORF did not replicate in protoplasts, while mutants in the initiation codon of the 1.9 kb ORF replicated at low levels. The two groups of mutants were not able to complement each other.

MeSH Terms
Brassica/genetics Cloning, Molecular Codon/genetics,physiology DNA/isolation & purification Mosaic Viruses/genetics Mutation Peptide Chain Initiation, Translational Plant Diseases RNA/genetics,physiology Transcription, Genetic
Chemicals
Codon RNA DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Weiland J J
Department of Agricultural Chemistry, Oregon State University, Corvallis 97331-6502.
Dreher T W
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/17.12.4675
Published
1989-06-26
Pages
4675-87
Language
English
Region
England
NLM ID
0411011
PMCID
PMC318024
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/17.12.4675 PMC Full Text View journal details
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