Cloning and characterization of two plasmids from Bacillus thuringiensis in Bacillus subtilis.

L Boe, Tim Tolker-Nielsen, S M Madsen, L Andrup, G Bolander

23 Citations (Scopus)

Abstract

Bacillus thuringiensis subspecies israliensis plasmids pTX14-1 and pTX14-3 were cloned and analyzed by Southern blot hybridization for their replication mechanism in Bacillus subtilis. The cloning of pTX14-1 into the replicon deficient vector pBOE335 showed the usual characteristics of single-stranded DNA plasmids, i.e., it generated circular single-stranded DNA and high molecular weight (HMW) multimers. The other plasmid, pTX14-3, behaved differently; it generated neither single-stranded DNA nor HMW multimers. Treatment with rifampicin did not result in the accumulation of single-stranded DNA. However, deletion of an EcoRI-PstI fragment resulted in the accumulation of both single-stranded DNA and HMW multimers. From various deletion derivatives, we have mapped the minus origin and the locus responsible for suppression of HMW multimer formation. Full activity of the minus origin and of the locus suppressing HMW formation was only observed on the native replicon, indicating a coupling to the plus strand synthesis.
Original languageEnglish
JournalPlasmid
Volume25
Issue number3
Pages (from-to)190-7
Number of pages7
ISSN0147-619X
Publication statusPublished - 1991
Externally publishedYes

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