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Molecular cloning and sequence analysis of the lysR gene from the extremely thermophilic eubacterium, Thermus thermophilus HB27
Authors:Takehide Kosuge  Takayuki Hoshino
Institution:Centre International en Recherche Agronomique pour le Développement, Département Elevage et Médecine Vétérinaire (CIRAD-EMVT), BP 515, 97165 Pointe àPitre Cedex, Guadeloupe;CIRAD-EMVT, BP 5035, 34032 Montpellier Cedex 1, France
Abstract:We have isolated a lysine-auxotrophic and kanamycin-resistant mutant from an extreme thermophile, Thermus thermophilus HB27. This mutant showed the lysA? or lysR? genotype since it could not grow on the minimal plate which contained diaminopimelic acid. Sequence analysis of the clones which could rescue the Lys? mutant indicated the lysR gene. The lysR gene overlapped with the rimK gene for the modification enzyme of ribosomal protein S6. In the Lys? mutant, the lysR gene was disrupted and the C-terminus region of the RimK protein was different from that of the wild-type, which contributed to the Lys? and kanamycin-resistant phenotype. The deduced amino acid sequence of the lysR gene showed 20.9% identity with the LysR protein of Escherichia coli. The percentage of use of cytosine or guanine in the third letter of the codons in the lysR gene was only 67.4%. We also determined that the argC gene encoding N-acetyl-γ-glutamyl phosphate reductase and the argB gene encoding acetylglutamate kinase were located immediately upstream of the lysR gene.
Keywords:Cowdria ruminantium            RAPD assay  Restriction fragment length polymorphism  Genetic diversity
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