Vectors permitting visual monitoring of simple transposition events.
The construction and use of two novel transposon(Tn)-delivery vectors is described. These vectors carry Inc.W or Inc.N broad-host-range transfer functions cloned next to the narrow-host-range replicon of pBR329. The host specificities of pSLX10 and pSLX23 both complement and extend the host specificities of existing Tn delivery vectors. Plasmids pSLX10 and pSLX23 were shown to transfer at high frequency in intergeneric matings. The lux genes which are present on each vector permit the visual monitoring of transconjugants which have retained a Tn element, but are devoid of plasmid molecules. pSLX10 and pLSX23 were efficiently used to generate a range of auxotrophic mutants in various strains of Pseudomonas as well as to clone genes from Serratia liquefaciens. These vectors may have general applicability to identify and clone genes in a wide range of Gram-negative bacteria.
Kozlowski
M
M
Allelix Biopharmaceuticals, Mississauga, Ont., Canada.
Wilkinson
D
D
Nash
N
N
Davies
R W
RW
D001426:Bacterial Proteins / Q000235:genetics*; D004251:DNA Transposable Elements; D004274:DNA, Recombinant / Q000032:analysis; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D008163:Luminescent Measurements; D010088:Oxidoreductases; D010957:Plasmids; D011549:Pseudomonas / Q000235:genetics; D012093:Replicon; D014733:Vibrio / Q000235:genetics
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004274:DNA, Recombinant; D010088:Oxidoreductases; C055198:LuxC protein, Bacteria