Desomer J, Vereecke D, Crespi M, Van Montagu M
Laboratorium voor Genetica, Universiteit Gent, Belgium.
Mol Microbiol. 1992 Aug;6(16):2377-85. doi: 10.1111/j.1365-2958.1992.tb01412.x.
The nucleotide sequence of the chloramphenicol-resistance gene (cmr) of Rhodococcus fascians NCPPB 1675 (located on the conjugative plasmid pRF2) allowed the identification of two possible open reading frames (ORFs), of which ORF1 was consistent with the mutational analysis. Biochemical analysis of cmr revealed that it does not encode an antibiotic-modifying enzyme. The amino acid sequence of ORF1 predicted a hydrophobic protein, with 12 putative membrane-spanning domains, homologous to proteins involved in the efflux of tetracycline across the plasma membrane. Expression of the cmr gene was induced by addition of chloramphenicol to the growth media. The promoter of this gene was restricted to 50 bp upstream from a 200 bp 5'-untranslated mRNA region, the latter containing two inverted repeats. At the amino acid level, the cmr gene is 52% identical to a previously identified chloramphenicol-resistance determinant in Streptomyces lividans, indicating a wider dispersion of this type of cmr gene among the actinomycetes.
法氏红球菌NCPPB 1675(位于接合质粒pRF2上)的氯霉素抗性基因(cmr)的核苷酸序列使得能够鉴定出两个可能的开放阅读框(ORF),其中ORF1与突变分析结果一致。对cmr的生化分析表明,它不编码抗生素修饰酶。ORF1的氨基酸序列预测为一种疏水蛋白,具有12个假定的跨膜结构域,与参与四环素跨质膜外排的蛋白同源。向生长培养基中添加氯霉素可诱导cmr基因的表达。该基因的启动子局限于200 bp 5'-非翻译mRNA区域上游的50 bp,后者包含两个反向重复序列。在氨基酸水平上,cmr基因与先前在淡紫链霉菌中鉴定出的氯霉素抗性决定簇有52%的同一性,表明这种类型的cmr基因在放线菌中分布更广泛。