Delivery system for creation of one-step in vivo lac gene fusions in Pseudomonas spp. involved in biological control.
The suicide plasmid pVA838 carrying the operon fusion transposon Tn5-lac was used as a delivery system to introduce Tn5-lac into Pseudomonas sp. strain M114. Random, in vivo lac gene fusions were successfully isolated in a one-step conjugation approach with this vector system. Tn5-lac-containing exconjugants were recovered at a frequency of approximately 10(-7) per recipient. However, when the mating temperature was increased from the normal growth temperature (28 degrees C) to 34 degrees C, the frequency was increased to approximately 10(-4) per recipient. A number of in vivo lac gene fusions were isolated and characterized in strain M114, a potentially important bacterium for biological control purposes.
O'Sullivan
D J
DJ
Microbiology Department, University College, Cork, Ireland.
O'Gara
F
F
D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D009154:Mutation; D010957:Plasmids; D011549:Pseudomonas / Q000235:genetics* / Q000254:growth & development; D013696:Temperature
D004251:DNA Transposable Elements