Tseng Chih-Sian, Yen Yu-Chieh, Chang Chao-Chin, Hsu Yuan-Man
Department of Biological Science and Technology, College of Life Sciences, China Medical University, Taichung, Taiwan.
Graduate Institute of Microbiology and Public Health, School of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan.
Biomedicine (Taipei). 2014;4(3):20. doi: 10.7603/s40681-014-0020-3. Epub 2014 Aug 13.
Integrons, mobile genetic units, capture and incorporate antibiotic resistance gene cassette by site-specific recombination. Class 1 integrons are widespread and associated with dispersion of antibiotic resistance among Gram-negative bacteria. The expression of gene cassette in Class 1 can vary, based on the Pc promoter but seldom from another promoter hiding downstream of Pc, called P. To probe distribution and prevalence of gene cassette promoter variants, we analyzed 169 . Choleraesuis and 191 . Typhimurium isolates from humans and animals, finding 95.27% occurrence of integrin among S. Choleraesuis, 83.25% among . Typhimurium. PCR-RFLP analysis identified four promoters (PcS+P, PcW+P, PcH1+P, and Pc+P-GGG) in said integron-positive isolates; major types in . Choleraesuis and . Typhimurium were PcS+P and Pc+P, respectively. Likewise, β-galactosidase assay rated promoter strength of variants by transcriptional fusion constructs to show extended -10 promoter (TGn/-10 promoter) in Pc and three-nucleotide insertion (GGG) between -35 and -10 region of P improving promoter strength of gene cassette.
整合子是可移动的遗传单位,通过位点特异性重组捕获并整合抗生素抗性基因盒。1类整合子广泛存在,并与革兰氏阴性菌中抗生素抗性的传播有关。1类基因盒的表达可能会有所不同,这取决于Pc启动子,但很少来自隐藏在Pc下游的另一个启动子P。为了探究基因盒启动子变体的分布和流行情况,我们分析了169株猪霍乱沙门氏菌和191株鼠伤寒沙门氏菌的人类和动物分离株,发现猪霍乱沙门氏菌中整合子的出现率为95.27%,鼠伤寒沙门氏菌中为83.25%。PCR-RFLP分析在上述整合子阳性分离株中鉴定出四种启动子(PcS+P、PcW+P、PcH1+P和Pc+P-GGG);猪霍乱沙门氏菌和鼠伤寒沙门氏菌中的主要类型分别为PcS+P和Pc+P。同样,β-半乳糖苷酶分析通过转录融合构建体评估变体的启动子强度,结果显示Pc中的延伸-10启动子(TGn/-10启动子)以及P的-35和-10区域之间的三核苷酸插入(GGG)提高了基因盒的启动子强度。