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Abstract
A large number of species of corynebacteria are known to be amino acid producers, including members of the genera Corynebacteriumand Brevibacterium.Pulsed-field gel electrophoresis (PFGE) of DNA fragments obtained by using endonucleases which recognize AT-rich hexanucleotide or octanucleotide sequences produces a discrete pattern of bands useful for fingerprinting and physical mapping of the chromosome. Using Pacland Swalendonucleases the genome of Brevibacterium lactofermentumATCC 13869 (genome size 3052 kb) was consistently cut into 26 and 20 bands, respectively, and the genome of Corynebacterium glutamicumATCC 13032 (2987 kb) yielded 27 and 26 fragments, respectively. The pattern of restriction fragments was identical for related strains (B. lactofermentumATCC 13869, B. lactofermentumBLO, B. lactofermentumR31) but different from the pattern of fragments of other soil isolates of the same species (B. lactofermentumDSM 20412) or from closely related organisms such as C. glutamicum; the different pattern of restriction fragments may be used to differentiate taxonomically related species. Brevibacterium linensshowed a different behaviour, due to its high G + C content; its genome (3105 kb) was resolved into 8 or 15 fragments, respectively, by digestion with the hexanucleotide-recognizing endonucleases Draland Asel.PFGE of DNA fragments obtained using these enzymes is a powerful technique for quick resolution of the corynebacteria genome into a small number of large fragments.
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