Open AccessConstruction of mini‐Tn 10luxABcam/Ptac ‐ATS and its use for developing a bacteriophage that transduces bioluminescence to Escherichia coli O157:H7
Author(s) -
Waddell Thomas E,
Poppe Cornelius
Publication year - 2000
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.1574-6968.2000.tb08909.x
Subject(s) - escherichia coli , bacteriophage , vibrio harveyi , biology , microbiology and biotechnology , bioluminescence , plasmid , prophage , lysogenic cycle , transposable element , luciferase , shuttle vector , virology , bacteria , vibrio , genetics , gene , mutant , vector (molecular biology) , biochemistry , recombinant dna , transfection
Mini‐Tn 10luxABcam/Ptac ‐ATS was constructed in order to develop a luciferase‐transducing bacteriophage for detecting Escherichia coli O157:H7. The transposon was designed to deliver a 3.6‐kb insertion that confers n ‐decanal‐dependent bioluminescence and resistance to chloramphenicol and was constructed using mini‐Tn 10cam/Ptac ‐ATS in the plasmid pNK2884 and luxAB from Vibrio harveyi . ΦV10, a temperate bacteriophage infecting common phage types of Escherichia coli O157:H7, was mutagenized as a prophage in E. coli O157:H7 strain R508. ΦV10:: luxABcam A1‐23 was rescued from the strain by propagating it on a strain lacking the bacteriophage and the vector containing the transposon. The bacteriophage transduced n ‐decanal‐dependent bioluminescence to E. coli O157:H7 strain R508 that was measurable approximately 1 h post infection.