Open AccessTransposon Tn5 mutagenesis in Rhodopseudomonas palustris
Author(s) -
Elder Douglas J.E.,
Morgan Philip,
Kelly David J.
Publication year - 1993
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.1574-6968.1993.tb06356.x
Subject(s) - rhodopseudomonas palustris , transposon mutagenesis , transposable element , biology , plasmid , rhodopseudomonas , mutagenesis , kanamycin , mutant , escherichia coli , microbiology and biotechnology , bacteria , genetics , dna , gene , antibiotics
The potential of the antibiotic resistance transposon Tn5 for random insertion mutagenesis in Rhodopseudomonas palustris was assessed. The Tn5 containing suicide vector plasmid pSUP2021, was transferred from Escherichia coli to Rhodopseudomonas palustris and kanamycin‐resistant transconjugants arose at a frequency of 2.7×10 −7 per recipient. In the majority of transconjugants tested, Tn5 was found to have successfully transposed to yield a single chromosomal insertion, with the concomitant loss of the vector plasmid through segregation. Two Tn5 mutants, one defective in carotenoid synthesis, and one exhibiting a reduced anaerobic growth rate on aromatic acids, were partially characterised. This is the first study to show that Tn5 mutagenesis can be applied successfully to isolate mutants of Rhodopseudomonas palustris .