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PMID: 3112515 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Mutations affecting translation of the bacteriophage T4 rIIB gene cloned in Escherichia coli.

Molecular & general genetics : MGG ·Vol. 207 ·No. 2-3 ·1987-05-00 ·Pages 224-32

Shinedling S, Gayle M, Pribnow D, Gold L

Abstract

Mutant ribosome binding sites of the bacteriophage T4 rIIB gene, resident on an 873 bp DNA fragment, were cloned into a plasmid vector as in-frame fusions to a reporter gene, beta-galactosidase. The collection of mutations included changes in the region 5' to the Shine/Dalgarno sequence, a mutation of the Shine/Dalgarno sequence, the alternate initiation codons GUG, AUA and ACG, and mutants in which several closely spaced initiation codons compete with each other on the same mRNA. The results show that the secondary structure variations we have installed 5' to the Shine/Dalgarno sequence have little effect on translation. GUG is essentially as good an initiator of translation as AUG when they are assayed on separate messages, but is outcompeted at least 50-fold in the sequence AUGUG. AUA and ACG are poor start codons, and are temperature sensitive. The initiation codon pair AUGAUA, in which the AUG is only two nucleotides from the Shine/Dalgarno sequence, displays a novel cold-sensitive phenotype.

MeSH Terms
Base Sequence Cloning, Molecular Escherichia coli/genetics Genes Genes, Viral Mutation Nucleic Acid Conformation Plasmids Protein Biosynthesis RNA, Messenger/genetics Ribosomes/metabolism T-Phages/genetics Viral Proteins/genetics beta-Galactosidase/genetics
Chemicals
RNA, Messenger Viral Proteins beta-Galactosidase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Shinedling S
Gayle M
Pribnow D
Gold L
References (36)
36 references, click to expand
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00331582
Published
1987-05-00
Pages
224-32
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
Grants
NIGMS NIH HHS · GM 28685 · United States
External Links
PubMed source DOI: 10.1007/BF00331582
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