A plasmid vector with a selectable marker for halophilic archaebacteria.
A mutant resistant to the gyrase inhibitor novobiocin was selected from a halophilic archaebacterium belonging to the genus Haloferax. Chromosomal DNA from this mutant was able to transform wild-type cells to novobiocin resistance, and these transformants formed visible colonies in 3 to 4 days on selective plates. The resistance gene was isolated on a 6.7-kilobase DNA KpnI fragment, which was inserted into a cryptic multicopy plasmid (pHK2) derived from the same host strain. The recombinant plasmid transformed wild-type cells at a high efficiency (greater than 10(6)/micrograms), was stably maintained, and could readily be reisolated from transformants. It could also transform Halobacterium volcanii and appears to be a useful system for genetic analysis in halophilic archaebacteria.
Holmes
M L
ML
Department of Microbiology, University of Melbourne, Parkville, Victoria, Australia.
Dyall-Smith
M L
ML
D001105:Archaea / Q000187:drug effects / Q000235:genetics*; D001419:Bacteria / Q000235:genetics*; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D004352:Drug Resistance, Microbial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006217:Halobacterium / Q000235:genetics*; D008969:Molecular Sequence Data; D009675:Novobiocin / Q000494:pharmacology; D010957:Plasmids; D012335:RNA, Ribosomal / Q000235:genetics / Q000302:isolation & purification; D015183:Restriction Mapping; D014169:Transformation, Bacterial
D004269:DNA, Bacterial; D012335:RNA, Ribosomal; D009675:Novobiocin