Identification of expression signals of the mycobacteriophages Bxb1, L1 and TM4 using the Escherichia-Mycobacterium shuttle plasmids pYUB75 and pYUB76 designed to create translational fusions to the lacZ gene

R. G. Barletta, D. D. Kim, S. B. Snapper, B. R. Bloom, W. R. Jacobs

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Mycobacterial expression signals were cloned using specially constructed gene fusion shuttle plasmid probes carrying a truncated Escherichia coli lacZ (β-galactosidase) gene which lacked a promoter, a ribosome binding site, and an ATG start codon. Libraries of mycobacteriophage Bxb1, L1 and TM4 DNAs were constructed, and introduced by electroporation into Mycobacterium smegmatis and the 'bacille Calmette-Guerin' (BCG). Clones carrying mycobacterial expression sequences were detected by their blue colour or characteristic fluorescence when plated on media containing chromogenic or fluorogenic substrates. Varying degrees of β-galactosidase expression were observed, and one Bxb1 expression signal was identified where β-galactosidase expression is repressed in phage lysogens.

Original languageEnglish (US)
Pages (from-to)23-30
Number of pages8
JournalJournal of General Microbiology
Volume138
Issue number1
DOIs
StatePublished - Jan 1 1992

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Mycobacteriophages
Galactosidases
Escherichia
Lac Operon
Mycobacterium
Plasmids
Mycobacterium smegmatis
Chromogenic Compounds
Electroporation
Initiator Codon
Gene Fusion
Ribosomes
Fluorescent Dyes
Bacteriophages
Clone Cells
Color
Fluorescence
Binding Sites
Escherichia coli
DNA

ASJC Scopus subject areas

  • Microbiology

Cite this

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AU - Kim, D. D.

AU - Snapper, S. B.

AU - Bloom, B. R.

AU - Jacobs, W. R.

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