Molecular cloning and sequence analysis of the lysR gene from the extremely thermophilic eubacterium, Thermus thermophilus HB27 |
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Authors: | Takehide Kosuge Takayuki Hoshino |
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Affiliation: | 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 |
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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. |
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Keywords: | Cowdria ruminantium RAPD assay Restriction fragment length polymorphism Genetic diversity |
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