Characterization of an insertion in the phage φ105 genome that blocks host Bacillus subtilis lysis and provides strong expression of heterologous genes

Yun Chung Leung, Jeffery Errington

Research output: Journal article publicationJournal articleAcademic researchpeer-review

16 Citations (Scopus)

Abstract

A defective prophage vector, φ105MU331, for high-level protein overproduction in Bacillus subtilis, was derived by random insertion of a lacZ reporter gene. The site of insertion not only provided efficient inducible transcription of heterologous genes, but also prevented lysis of the host cell. The region of the insertion in φ105MU331 lies close to the right cohesive end of φ105. DNA sequence analysis revealed that this region of φ105 somewhat resembles the lysis cassette of various phages, including λ. The site of insertion lies in a possible 'holin' gene, which could explain the block in host cell lysis. Dual promoters apparently responsible for the strong inducible transcription lie in an untranslated region just upstream from the putative holin gene. This region is probably equivalent to the site of the major late promoter and antiterminator of the lambdoid phages. The sequence features could, thus, account for the useful properties of the φ105MU331 vector system.
Original languageEnglish
Pages (from-to)1-6
Number of pages6
JournalGene
Volume154
Issue number1
DOIs
Publication statusPublished - 27 Feb 1995
Externally publishedYes

Keywords

  • defective prophage
  • DNA sequencing
  • dual promoters
  • dual start motif
  • holin gene
  • inducible expression
  • lacZ reporter gene
  • Lysis gene cluster
  • phage λ
  • transcription

ASJC Scopus subject areas

  • Genetics

Cite this