Artificial regulation of gene expression in Escherichia coli by RNase P

Cecilia Guerrier-Takada, Ying Li, Sidney Altman

Research output: Contribution to journalArticlepeer-review

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
Externally publishedYes

Keywords

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

ASJC Scopus subject areas

  • General

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