Determination of the nucleotide sequence for the glutamate synthase structural genes of Escherichia coli K-12

Guillermo Oliver, Guillermo Gosset, Ray Sanchez-Pescador, Edmundo Lozoya, Lailig M. Ku, Noemi Flores, Baltazar Becerril, Fernando Valle, Francisco Bolivar*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

58 Scopus citations

Abstract

We have determined the complete nucleotide sequence of a 6.3-kb chromosomal HpaI-EcoRI fragment, that contains the structural genes for both the large and small subunits of the Escherichia coli K-12 glutamate synthase (GOGAT) enzyme, as well as the 5 ' - and 3 ' -flanking and intercistronic DNA regions. The Mrs of the two subunits, as deduced from the nucleotide (nt) sequence, were estimated as 166 208 and 52 246. Partial amino acid sequence of the GOGAT enzyme revealed that the large subunit starts with a cysteine residue that is probably generated by a proteolytic cleavage. Northern blotting experiments revealed a transcript of approximately 7300 nt, that at least contains the cistrons for both subunits. A transcriptional start point and a functional promoter were identified in the 5' DNA flanking region of the large subunit gene. The messenger RNA nontranslated leader region has 120 nt and shares identity with the leader regions of E. coli ribosomal operons, in particular around the so-called boxA sequence implicated in antitermination. Other possible regulatory sequences are described.

Original languageEnglish (US)
Pages (from-to)1-11
Number of pages11
JournalGene
Volume60
Issue number1
DOIs
StatePublished - 1987

Keywords

  • Recombinant DNA
  • flavoprotein
  • functional promoter
  • nitrogen metabolism
  • oligodeoxynucleotide primer
  • protein amino terminus
  • transcriptional start point

ASJC Scopus subject areas

  • Genetics

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