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New thermosensitive plasmid for gram-positive bacteria.

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  • 1. Laboratoire de Génétique Microbienne, Institut National de la Recherche Agronomique-Domaine de Vilvert, Jouy en Josas, France.
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    • Maguin E 1
    (1 author)

ORCIDs linked to this article

Journal of Bacteriology, 01 Sep 1992, 174(17):5633-5638
https://doi.org/10.1128/jb.174.17.5633-5638.1992 PMID: 1324906 PMCID: PMC206509

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Abstract 

We isolated a replication-thermosensitive mutant of the broad-host-range replicon pWV01. The mutant pVE6002 is fully thermosensitive above 35 degrees C in both gram-negative and gram-positive bacteria. Four clustered mutations were identified in the gene encoding the replication protein of pVE6002. The thermosensitive derivative of the related plasmid pE194 carries a mutation in the analogous region but not in the same position. Derivatives of the thermosensitive plasmid convenient for cloning purposes have been constructed. The low shut-off temperature of pVE6002 makes it a useful suicide vector for bacteria which are limited in their own temperature growth range. Using pVE6002 as the delivery vector for a transposon Tn10 derivative in Bacillus subtilis, we observed transposition frequencies of about 1%.

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J Bacteriol. 1992 Sep; 174(17): 5633–5638.
PMCID: PMC206509
PMID: 1324906

New thermosensitive plasmid for gram-positive bacteria.

E Maguin, P Duwat, T Hege, D Ehrlich, and A Gruss
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Abstract

We isolated a replication-thermosensitive mutant of the broad-host-range replicon pWV01. The mutant pVE6002 is fully thermosensitive above 35 degrees C in both gram-negative and gram-positive bacteria. Four clustered mutations were identified in the gene encoding the replication protein of pVE6002. The thermosensitive derivative of the related plasmid pE194 carries a mutation in the analogous region but not in the same position. Derivatives of the thermosensitive plasmid convenient for cloning purposes have been constructed. The low shut-off temperature of pVE6002 makes it a useful suicide vector for bacteria which are limited in their own temperature growth range. Using pVE6002 as the delivery vector for a transposon Tn10 derivative in Bacillus subtilis, we observed transposition frequencies of about 1%.

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Selected References

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