A chromosomal genetic map of Rhizobium sp. NGR234 generated with Tn5-Mob |
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Authors: | Magne ?ster?s John Stanley William J Broughton and David N Dowling |
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Institution: | (1) L.B.M.P.S., Université de Genève, 1, chemin de l'Impératrice, 1292 Chambésy, Geneve, Switzerland;(2) Bereich Mikrobiologie, Gesellschaft für Biotechnologische Forschung (GBF), Mascheroder Weg 1, D-3300 Braunschweig, Federal Republic of Germany |
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Abstract: | Summary
Rhizobium sp. NGR234 in a fast-growing Rhizobium strain with a broad host range. The location and role of chromosomal genes involved in cellular metabolism or in the legume symbioses is unknown. We isolated a series of auxotrophic and antibiotic resistant mutants of NGR234 and utilized a chromosome mobilization system based on Tn5-Mob and pJB3JI; Tn5-Mob donor strains behaved like Hfr strains, transferring the chromosome polarly at high frequency from a fixed point of insertion. The use of four different strains with Tn5-Mob located at different nutritional loci in crosses with double auxotrophic recipients, allowed us to build up a circular linkage map of NGR234 based on relative recombination frequencies. Also, symbiotically important genes identified by site-directed mutagenesis, such as hemA and ntrA, could be located and mapped on the chromosome.Abbreviations Tc
tetracycline
- Sp
spectinomycin
- Rif
rifampicin
- Km
kanamycin |
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Keywords: | Wide host-range Rhizobium Auxotrophic mutants Site-directed mutants Chromosome mobilization Linkage map |
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