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PMID: 3003520 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cloning of the Escherichia coli gene for the stringent starvation protein.

Molecular & general genetics : MGG ·Vol. 201 ·No. 2 ·1985-00-00 ·Pages 151-7

Fukuda R, Yano R, Fukui T, Hase T, Ishihama A, Matsubara H

Abstract

In order to clone the Escherichia coli gene for the stringent starvation protein (SSP), we determined its N-terminal sequence as well as the sequence of two peptide fragments obtained by cyanogen bromide cleavage of the protein. We then chemically synthesized four sets of oligodeoxyribonucleotide mixtures that represented possible codon combinations for parts of these amino acid sequences. The synthetic oligonucleotides were labelled with 32P at their 5'-termini and used as hybridization probes to detect DNA fragments containing the complementary sequences. Genomic Southern hybridization of E. coli chromosomal DNA gave up to ten DNA fragments hybridizing with each probe but only a few hybridized with two or more of the probes. The latter fragments were cloned in pBR322. By determining partial base sequences with a rapid method and examining proteins encoded by the DNA fragments, we were able to show that we had isolated a clone containing the complete SSP structural gene.

MeSH Terms
Amino Acid Sequence Base Sequence Cloning, Molecular DNA Restriction Enzymes DNA, Superhelical/genetics Escherichia coli/genetics GTP Pyrophosphokinase/genetics Genes Genes, Bacterial Nucleic Acid Hybridization Phosphotransferases/genetics Plasmids
Chemicals
DNA, Superhelical Phosphotransferases GTP Pyrophosphokinase DNA Restriction Enzymes
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Fukuda R
Yano R
Fukui T
Hase T
Ishihama A
Matsubara H
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26 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/BF00425652
Published
1985-00-00
Pages
151-7
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00425652
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