Open AccessLow target site specificity of an IS 6100 ‐based mini‐transposon, Tn 1792 , developed for transposon mutagenesis of antibiotic‐producing Streptomyces
Author(s) -
Herron Paul R,
Evans Meirwyn C,
Dyson Paul J
Publication year - 1999
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.1574-6968.1999.tb13435.x
Subject(s) - transposon mutagenesis , transposable element , streptomyces , biology , genetics , sleeping beauty transposon system , transposition (logic) , mutagenesis , plasmid , tn3 transposon , genome , gene , mutation , bacteria , linguistics , philosophy
To improve transposon mutagenesis of antibiotic‐producing Streptomyces , a mini‐transposon, Tn 1792 , was constructed, based on IS 6100 , originally isolated from Mycobacterium fortuitum . Easily manageable transposition assays were developed to demonstrate inducible transposition of Tn 1792 into the Streptomyces genome from a temperature‐sensitive delivery plasmid. Introduction of the selectable aac1 gene between the inverted repeats in Tn 1792 allowed for both reliable identification of transposition events in Streptomyces , and also subsequent cloning of transposon‐tagged sequences in Escherichia coli . This enabled the target site specificity of Tn 1792 to be determined at nucleotide resolution, revealing no significant shared homology between different target sites. Consequently, Tn 1792 is well suited for random mutagenesis of Streptomyces .