β-Lactamase expression in Streptomyces cacaoi

H. Urabe, M. V. Lenzini, M. Mukaide, J. Dusart, M. M. Nakano, J. M. Ghuysen, H. Ogawara

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Plasmids were prepared by inserting genomic DNA fragments from Streptomyces cacaoi within the mel gene of plasmid pIJ702. The inserted DNA fragments contain the β-lactamase-encoding bla gene and upstream nucleotide sequences of various lengths. The transcription start point of bla was identified by nuclease S1 mapping. Upstream nucleotide sequences of sufficient lengths had an enhancing effect on β-lactamase production by the Steptomyces host. The dot blot hybridization assay revealed that this effect was exerted at the transcriptional level. Experimental evidence strongly suggests that the underlying mechanism involves, at least in part, one or several trans-acting elements. In one of the constructs, in which the upstream nucleotide sequence was reduced to 0.3 kb, the bla promoter was present but the bla gene was expressed by readthrough from a promoter, possibly the mel promoter, of the pIJ702 vector.

Original languageEnglish (US)
Pages (from-to)6427-6434
Number of pages8
JournalJournal of bacteriology
Volume172
Issue number11
DOIs
StatePublished - 1990

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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