Enhancement of translation initiation by A/T-rich sequences downstream of the initiation codon in Escherichia coli

Guoliang Qing, Bing Xia, Masayori Inouye

Research output: Contribution to journalArticlepeer-review

50 Scopus citations

Abstract

The region located downstream of the initiation codon constitutes part of the translation initiation signal, significantly affecting the level of protein expression in E. coli. In order to determine its influence on translation initiation, we inserted random 12-base sequences downstream of the initiation codon of the lacZ gene. A total of 119 random clones showing higher β-galactosidase activities than the control lacZ gene were isolated and subsequently sequenced. Analysis of these clones revealed that their insertion sequences are strikingly rich in A and T, but poor in G, with no consensus sequences among them. Toeprinting experiments and polysome profile analysis confirmed that the A/T-rich sequences enhance translation at the level of initiation. Collectively, the present data demonstrate that A/T richness of the region following the initiation codon plays a significant role in E. coli gene expression.

Original languageEnglish (US)
Pages (from-to)133-144
Number of pages12
JournalJournal of molecular microbiology and biotechnology
Volume6
Issue number3-4
DOIs
StatePublished - 2003

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Microbiology
  • Applied Microbiology and Biotechnology
  • Molecular Biology

Keywords

  • Downstream A/T-rich sequence
  • Escherichia coli
  • Toeprinting
  • Translation initiation enhancement
  • lacZ

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