Direct selection of recombinant plasmids in Bacillus subtilis

Thomas J. Gryczan, David Dubnau

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

A system is described which permits the direct, positive selection of recombinant plasmids in Bacillus subtilis. This system relies on the plasmid pBD214 which confers chloramphenicol (Cm) resistance and carries a thy gene, and on BD393, a highly competent B. subtilis thy A thy B host. Thy- strains are resistant to trimethoprim (Tmp), and Thy+ strains are sensitive. Inactivation of the pBD214 thy determinant by insertion of a DNA fragment permits selection of Cmr Tmpr clones, all of which carry recombinant plasmids. This insertional inactivation can be accomplished using the unique EcoRl, Bell, Pvull, or EcoRV sites, all of which are located within the thy gene on pBD214. Some properties of this selective system are described, and its uses for molecular cloning are discussed.

Original languageEnglish (US)
Pages (from-to)459-469
Number of pages11
JournalGene
Volume20
Issue number3
DOIs
StatePublished - Dec 1982

All Science Journal Classification (ASJC) codes

  • Genetics

Keywords

  • B. subtilis cloning
  • Recombinant DNA
  • gene libraries
  • plasmid vector
  • thy inactivation
  • trimethoprim resistance

Fingerprint

Dive into the research topics of 'Direct selection of recombinant plasmids in Bacillus subtilis'. Together they form a unique fingerprint.

Cite this