Open AccessConstruction and use of a new vector/transposon, pLBT::mim‐Tn 10:lac:kan , to identify environmentally responsive genes in a marine bacterium
Author(s) -
Albertson Nan H.,
Stretton Serina,
Pongpattanakitshote Somchai,
Östling Jörgen,
Marshall Kevin C.,
Goodman Amanda E.,
Kjelleberg Staffan
Publication year - 1996
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1996.tb08351.x
Subject(s) - transposable element , tn10 , biology , plasmid , mutant , gene , reporter gene , genetics , gene cassette , shuttle vector , transposon mutagenesis , sleeping beauty transposon system , vector (molecular biology) , bacteria , dna transposable elements , gene expression , integron , recombinant dna
The previously described pLOFKm transposon delivery plasmid (J. Bacteriol. (1990) 172, 6557–6567) was engineered such that a promoterless lacZ gene was cloned within the transposon cassette, generating the vector pLBT. Using pLBT, stable insertion mutations were generated at high frequencies in Vibrio sp. S141 and Pseudomonas sp. S91, and the interrupted genes could be monitored for their pattern of regulation. Genetic screens isolated mutants defective in a variety of activities. We describe the construction and use of pLBT as a tool for reporter gene mutant analysis in bacteria other than well‐characterized laboratory strains.