Transcription of single-copy hybrid lacZ genes by T7 RNA polymerase in Escherichia coli: mRNA synthesis and degradation can be uncoupled from translation.

Abstract:

:In Escherichia coli transcription of individual genes generally requires concomitant translation, and thus the decay of mRNAs cannot be studied without the complication of translation. Here we have used T7 RNA polymerase to transcribe in vivo lacZ genes carrying ribosome binding sites of variable efficiency. We show that neither cell viability nor growth rate is affected by the T7-driven transcription of these genes, provided that they are present as single chromosomal copy. Furthermore, transcription is now completely uncoupled from translation, allowing large amounts of even completely untranslated mRNAs to be synthesized. Taking advantage of these features, we discuss the influence of the frequency of translation upon the processing and degradation of the lac message.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Chevrier-Miller M,Jacques N,Raibaud O,Dreyfus M

doi

10.1093/nar/18.19.5787

subject

Has Abstract

pub_date

1990-10-11 00:00:00

pages

5787-92

issue

19

eissn

0305-1048

issn

1362-4962

journal_volume

18

pub_type

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