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嗜麦芽窄食单胞菌染色体aac(6')-Iz基因的特性分析

Characterization of the chromosomal aac(6')-Iz gene of Stenotrophomonas maltophilia.

作者信息

Lambert T, Ploy M C, Denis F, Courvalin P

机构信息

Unité des Agents Antibactériens, Institut Pasteur, 75724 Paris Cedex 15, France.

出版信息

Antimicrob Agents Chemother. 1999 Oct;43(10):2366-71. doi: 10.1128/AAC.43.10.2366.

Abstract

The aac(6')-Iz gene of Stenotrophomonas maltophilia BM2690 encoding an aminoglycoside 6'-N-acetyltransferase was characterized. The gene was identified as a coding sequence of 462 bp corresponding to a protein with a calculated mass of 16,506 Da, a value in good agreement with that of ca. 16,000 found by in vitro coupled transcription-translation. Analysis of the deduced amino acid sequence indicated that the protein was a member of the major subfamily of aminoglycoside 6'-N-acetyltransferases. The enzyme conferred resistance to amikacin but not to gentamicin, indicating that it was an AAC(6') of type I. The open reading frame upstream from the aac(6')-Iz gene was homologous to the fprA gene of Myxococcus xanthus (61% identity), which encodes a putative pyridoxine (pyridoxamine) 5'-phosphate oxidase. Pulsed-field gel electrophoresis of total DNA from BM2690 and S. maltophilia ATTC 13637 digested with XbaI, DraI, and SpeI followed by hybridization with rRNA and aac(6')-Iz-specific probes indicated that the gene was located in the chromosome. The aac(6')-Iz gene was detected by DNA-DNA hybridization in all 80 strains of S. maltophilia tested. The MICs of gentamicin against these strains of S. maltophilia were lower than those of amikacin, netilmicin, and tobramycin, indicating that production of AAC(6')-Iz contributes to aminoglycoside resistance in S. maltophilia.

摘要

对嗜麦芽窄食单胞菌BM2690中编码氨基糖苷6'-N-乙酰基转移酶的aac(6')-Iz基因进行了表征。该基因被鉴定为一个462 bp的编码序列,对应于一个计算分子量为16,506 Da的蛋白质,该值与通过体外偶联转录-翻译测得的约16,000 Da的值高度一致。对推导的氨基酸序列分析表明,该蛋白质是氨基糖苷6'-N-乙酰基转移酶主要亚家族的成员。该酶赋予对阿米卡星的抗性,但对庆大霉素无抗性,表明它是I型的AAC(6')。aac(6')-Iz基因上游的开放阅读框与黄色黏球菌的fprA基因同源(同一性为61%),fprA基因编码一种假定的吡哆醇(吡哆胺)5'-磷酸氧化酶。用XbaI、DraI和SpeI消化嗜麦芽窄食单胞菌BM2690和嗜麦芽窄食单胞菌ATCC 13637的总DNA,然后用rRNA和aac(6')-Iz特异性探针进行杂交,结果表明该基因位于染色体上。通过DNA-DNA杂交在所有测试的80株嗜麦芽窄食单胞菌中均检测到aac(6')-Iz基因。庆大霉素对这些嗜麦芽窄食单胞菌菌株的最低抑菌浓度低于阿米卡星、奈替米星和妥布霉素,表明AAC(6')-Iz的产生有助于嗜麦芽窄食单胞菌对氨基糖苷类抗生素产生抗性。

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