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PMID: 2668891 Published · ppublish English Comparative Study Journal Article

Isolation and characterization of the gene coding for Escherichia coli arginyl-tRNA synthetase.

Nucleic acids research ·Vol. 17 ·No. 14 ·1989-07-25 ·Pages 5725-36

Eriani G, Dirheimer G, Gangloff J

Abstract

The gene coding for Escherichia coli arginyl-tRNA synthetase (argS) was isolated as a fragment of 2.4 kb after analysis and subcloning of recombinant plasmids from the Clarke and Carbon library. The clone bearing the gene overproduces arginyl-tRNA synthetase by a factor 100. This means that the enzyme represents more than 20% of the cellular total protein content. Sequencing revealed that the fragment contains a unique open reading frame of 1734 bp flanked at its 5' and 3' ends respectively by 247 bp and 397 bp. The length of the corresponding protein (577 aa) is well consistent with earlier Mr determination (about 70 kd). Primer extension analysis of the ArgRS mRNA by reverse transcriptase, located its 5' end respectively at 8 and 30 nucleotides downstream of a TATA and a TTGAC like element (CTGAC) and 60 nucleotides upstream of the unusual translation initiation codon GUG; nuclease S1 analysis located the 3'-end at 48 bp downstream of the translation termination codon. argS has a codon usage pattern typical for highly expressed E. coli genes. With the exception of the presence of a HVGH sequence similar to the HIGH consensus element, ArgRS has no relevant sequence homologies with other aminoacyl-tRNA synthetases.

MeSH Terms
Amino Acid Sequence Amino Acyl-tRNA Synthetases/genetics Arginine-tRNA Ligase/genetics,isolation & purification Base Sequence Cloning, Molecular Escherichia coli/enzymology,genetics Genes Genes, Bacterial Molecular Sequence Data Plasmids RNA, Messenger/genetics Restriction Mapping Sequence Homology, Nucleic Acid Transcription, Genetic
Chemicals
RNA, Messenger Amino Acyl-tRNA Synthetases Arginine-tRNA Ligase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Eriani G
Institut de Biologie Moléculaire et Cellulaire du CNRS, Strasbourg, France.
Dirheimer G
Gangloff J
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/17.14.5725
Published
1989-07-25
Pages
5725-36
Language
English
Region
England
NLM ID
0411011
PMCID
PMC318192
Subset
IM
Databases
GENBANK
X15320
External Links
PubMed source DOI: 10.1093/nar/17.14.5725 PMC Full Text View journal details
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