School of Biological Sciences, University of Wollongong, Wollongong, New South Wales, Australia.
PLoS One. 2010 Sep 15;5(9):e12754. doi: 10.1371/journal.pone.0012754.
Class 1 integrons play a role in the emergence of multi-resistant bacteria by facilitating the recruitment of gene cassettes encoding antibiotic resistance genes. 512 E. coli strains sourced from humans (n = 202), animals (n = 304) and the environment (n = 6) were screened for the presence of the intI1 gene. In 31/79 integron positive E. coli strains, the gene cassette regions could not be PCR amplified using standard primers. DNA sequence analysis of 6 serologically diverse strains revealed atypical integrons harboured the dfrA5 cassette gene and only 24 bp of the integron 3'-conserved segment (CS) remained, due to the insertion of IS26. PCR targeting intI1 and IS26 followed by restriction fragment length polymorphism (RFLP) analysis identified the integron-dfrA5-IS26 element in 27 E. coli strains of bovine origin and 4 strains of human origin. Southern hybridization and transformation studies revealed the integron-dfrA5-IS26 gene arrangement was either chromosomally located or plasmid borne. Plasmid location in 4/9 E. coli strains and PCR linkage of Tn21 transposition genes with the intI1 gene in 20/31 strains, suggests this element is readily disseminated by horizontal transfer.
Class 1 整合子通过促进编码抗生素抗性基因的基因盒的募集,在多抗性细菌的出现中发挥作用。从人类(n=202)、动物(n=304)和环境(n=6)中筛选了 512 株大肠杆菌,以检测 intI1 基因的存在。在 31/79 个整合子阳性大肠杆菌菌株中,使用标准引物无法扩增基因盒区域。对 6 个血清学多样化的菌株进行 DNA 序列分析表明,异常整合子携带 dfrA5 盒基因,由于 IS26 的插入,整合子 3'-保守片段(CS)仅剩下 24 bp。针对 intI1 和 IS26 的 PCR 靶向和限制性片段长度多态性(RFLP)分析鉴定了 27 株牛源大肠杆菌菌株和 4 株人源大肠杆菌菌株中的整合子-dfrA5-IS26 元件。Southern 杂交和转化研究表明,整合子-dfrA5-IS26 基因排列要么位于染色体上,要么位于质粒上。4/9 株大肠杆菌中的质粒定位和 20/31 株中的 Tn21 转位基因与 intI1 基因的 PCR 连锁表明,该元件可通过水平转移轻易传播。