Inducibility of the Streptomyces traRts107-Ptra Expression Cassette in Mycobacterium smegmatis

Amanda Lim, Calvin Boon, Thomas Dick

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

An inducible Streptomyces expression cassette utilising the Ptra promoter and a temperature-sensitive allele of the TraR repressor from S. nigrifaciens plasmid pSN22 was tested in Mycobacterium smegmatis. Using reporter assays and Northern blot analysis, a marked increase of Ptra-directed transcription was observed upon a temperature shift from 28 to 37°C. These results show that the S. nigrifaciens promoterrepressor cassette is functional in M. smegmatis. However, comparison of the level of induced Ptra-directed transcription with the level of transcription directed by the strong mycobacterial promoter Phsp60 indicated that the relative strength of the Ptra promoter was low. Considering the severe limitation of inducible expression systems for mycobacteria, this Streptomyces cassette might be a useful starting point for the development of a compact and fully portable inducible mycobacterial expression cassette.

Original languageEnglish (US)
Pages (from-to)517-519
Number of pages3
JournalBiological Chemistry
Volume381
Issue number5-6
DOIs
StatePublished - Jan 1 2000

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Mycobacterium smegmatis
Streptomyces
Transcription
Temperature
Mycobacterium
Northern Blotting
Plasmids
Alleles
Assays

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Biochemistry
  • Clinical Biochemistry

Cite this

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abstract = "An inducible Streptomyces expression cassette utilising the Ptra promoter and a temperature-sensitive allele of the TraR repressor from S. nigrifaciens plasmid pSN22 was tested in Mycobacterium smegmatis. Using reporter assays and Northern blot analysis, a marked increase of Ptra-directed transcription was observed upon a temperature shift from 28 to 37°C. These results show that the S. nigrifaciens promoterrepressor cassette is functional in M. smegmatis. However, comparison of the level of induced Ptra-directed transcription with the level of transcription directed by the strong mycobacterial promoter Phsp60 indicated that the relative strength of the Ptra promoter was low. Considering the severe limitation of inducible expression systems for mycobacteria, this Streptomyces cassette might be a useful starting point for the development of a compact and fully portable inducible mycobacterial expression cassette.",
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Inducibility of the Streptomyces traRts107-Ptra Expression Cassette in Mycobacterium smegmatis. / Lim, Amanda; Boon, Calvin; Dick, Thomas.

In: Biological Chemistry, Vol. 381, No. 5-6, 01.01.2000, p. 517-519.

Research output: Contribution to journalArticle

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