Cloning of the rec-2 locus of Haemophilus influenzae.
A collection of transposon mutants of Haemophilus influenzae was constructed by additive transformation with mutagenized chromosomal DNA. A rec-2::miniTn10 km mutation was cloned from a transformation-defective member of the mutant collection, followed by the reconstruction of the wild-type rec-2 locus by recombination to create pDM62. Southern blots showed that the commonly studied Rec-2 mutant, Rd(DB117)rec-, contained either a large deletion or a substitution that removed part of rec-2 locus. A collection of transposon mutations in pDM62 was used to characterize the rec-2 locus by complementation. A corresponding collection of mutants was also constructed. A single segment was required to complement the transformation defect in Rd(DB117)rec-. All of the transformation-defective transposon mutants failed to translocate donor DNA into then cell, in agreement with previous studies of Rd(DB117)rec-.
McCarthy
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Department of Botany and Microbiology, University of Oklahoma, Norman 73019.
D015139:Blotting, Southern; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D005798:Genes, Bacterial; D006193:Haemophilus influenzae / Q000235:genetics*; D009154:Mutation; D010957:Plasmids; D011995:Recombination, Genetic; D014170:Transformation, Genetic
D004251:DNA Transposable Elements