New thermosensitive plasmid for gram-positive bacteria.
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%.
Maguin
E
E
Laboratoire de Génétique Microbienne, Institut National de la Recherche Agronomique-Domaine de Vilvert, Jouy en Josas, France.
Duwat
P
P
Hege
T
T
Ehrlich
D
D
Gruss
A
A
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D004265:DNA Helicases; D004261:DNA Replication; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D004268:DNA-Binding Proteins; D006094:Gram-Positive Bacteria / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids; D011506:Proteins; D015183:Restriction Mapping; D016415:Sequence Alignment; D013696:Temperature; D015534:Trans-Activators
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D011506:Proteins; D015534:Trans-Activators; C045264:replication initiator protein; D004265:DNA Helicases