A general method for the transfer of plasmid-borne mutant alleles to the chromosome of Klebsiella pneumoniae using bacteriophage λ : Transfer of pqq genes
✍ Scribed by Meulenberg, J. J. M. ;Loenen, W. A. M. ;Sellink, E. ;Postma, P. W.
- Publisher
- Springer
- Year
- 1990
- Tongue
- English
- Weight
- 364 KB
- Volume
- 220
- Category
- Article
- ISSN
- 0026-8925
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✦ Synopsis
A generally applicable method is described for reintroduction of mutant plasmid-borne alleles to the chromosome of Klebsiella pneumoniae using bacteriophage 2. We,used this method to make stable chromosomal transposon insertions in genes for biosynthesis of pyrroloquinoline quinone in K. pneumoniae Key words: Transposon mutations -Gene transfer -Recombinationpqq genes -KlebsieIla pneumoniae Klebsiella pneumoniae contains a membrane-bound glucose dehydrogenase (GLD) which uses pyrroloquinoline quinone (PQQ) as a cofactor (Neijssel et al. 1983). We are studying the genes involved in the biosynthesis of PQQ, but progress has been hampered by problems in selecting mutations in the chromosomal pqq genes and the inability to transfer mutations from one Klebsiella strain to another. We recently isolated a clone from a cosmid library of K. pneumon- iae, complementing a pqq mutation. We decided to mutagenise this clone by transposon insertion and to transfer the mutated region back into the chromosome, thus creating new and stable chromosomal pqq mutations.