Artificial regulation of gene expression in Escherichia coli by RNase P

Cecilia Guerrier-Takada, Ying Li, Sidney Altman

Research output: Contribution to journalArticle

83 Scopus citations

Abstract

Plasmids encoding various external guide sequences (EGSs) were constructed and inserted into Escherichia coli. In strains harboring the appropriate plasmids, the expression of fully induced β-galactosidase and alkaline phosphatase activity was reduced by more than 50%, while no reduction in such activity was observed in strains with nonspecific EGSs. The inhibition of gene expression was virtually abolished at restrictive temperatures in strains that were temperature-sensitive for RNase P (EC 3.1.26.5). Northern blot analysis showed that the steady-state copy number of EGS RNAs was several hundred per cell in vivo. A plasmid that contained a gene for M1 RNA covalently linked to a specific EGS reduced the level of expression of a suppressor tRNA that was encoded by a separate plasmid. Similar methods can be used to regulate gene expression in E. coli and to mimic the properties of cold-sensitive mutants.

Original languageEnglish (US)
Pages (from-to)11115-11119
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume92
Issue number24
DOIs
StatePublished - Nov 21 1995

    Fingerprint

Keywords

  • M1 RNA
  • alkaline phosphatase
  • external guide sequences
  • suppressor tRNA
  • β-galactosidase

ASJC Scopus subject areas

  • General

Cite this