Centrón Daniela, Roy Paul H
Département de Biochimie et de Microbiologie, Faculté des Sciences et de Génie, Université Laval, Sainte-Foy, Québec, Canada G1K 7P4.
Antimicrob Agents Chemother. 2002 May;46(5):1402-9. doi: 10.1128/AAC.46.5.1402-1409.2002.
We analyzed the role of integrons in the dissemination of antibiotic resistance in a recent multiresistant clinical isolate, Serratia marcescens SCH88050909 (SCH909). This isolate harbors three integrons, all on a 60-kb conjugative plasmid. By PCR, hybridization, and sequencing analyses, we found that integron 1 has the dfrA1 and ant(3")-Ia cassettes. The first cassette in integron 2 contains the ant(2")-Ia gene, separated from its attC site (59-base element) by a 1,971-bp insert containing a group II intron; this intron codes for a putative maturase-reverse transcriptase on the complementary strand and is the first such intron to be found associated with an integron. The attC site is followed by a novel aminoglycoside resistance gene, ant(3")-Ii-aac(6')-IId, which has been characterized for its bifunctional ANT(3")-I and AAC(6')-II activities. DNA sequence analysis of this fused cassette suggests that insertion and excision due to the integrase activity could have an important role in the evolution of aminoglycoside resistance genes. This gene is followed by an unknown open reading frame with a typical attC site and a partial cassette composed of the beginning of the bla(OXA-10) cassette interrupted by IS1. The sequence downstream of IS1 revealed that the bla(OXA-10) cassette is incomplete and that the 3' conserved segment of this integron is absent. Integron 3 is in a Tn1696-like transposon with the aac(3)-Ia cassette followed by three unknown cassettes and ant(3")-Ia. The presence of the group II intron and the relationship of group II introns in eubacteria with mobile elements suggest a possible role of this element in events such as cassette formation and/or plasmid evolution.
我们分析了整合子在近期一株多重耐药临床分离株粘质沙雷氏菌SCH88050909(SCH909)抗生素耐药性传播中的作用。该分离株含有三个整合子,均位于一个60 kb的接合质粒上。通过聚合酶链反应(PCR)、杂交和测序分析,我们发现整合子1含有dfrA1和ant(3")-Ia基因盒。整合子2中的第一个基因盒包含ant(2")-Ia基因,该基因与其attC位点(59碱基元件)被一个1971 bp的插入片段隔开,该插入片段含有一个II类内含子;该内含子在互补链上编码一个推定的成熟酶-逆转录酶,是首次发现与整合子相关的此类内含子。attC位点后接一个新的氨基糖苷类耐药基因ant(3")-Ii-aac(6')-IId,该基因因其双功能ANT(3")-I和AAC(6')-II活性而得到表征。对这个融合基因盒的DNA序列分析表明,整合酶活性引起的插入和切除可能在氨基糖苷类耐药基因的进化中起重要作用。该基因后接一个具有典型attC位点的未知开放阅读框和一个由bla(OXA-10)基因盒开头被IS1中断组成的部分基因盒。IS1下游的序列显示bla(OXA-10)基因盒不完整,且该整合子的3'保守区段缺失。整合子3存在于一个Tn1696样转座子中,带有aac(3)-Ia基因盒,其后是三个未知基因盒和ant(3")-Ia。II类内含子的存在以及真细菌中II类内含子与可移动元件的关系表明,该元件可能在基因盒形成和/或质粒进化等事件中发挥作用。