Pulkkinen L, Huovinen P, Vuorio E, Toivanen P
Antimicrob Agents Chemother. 1984 Jul;26(1):82-6. doi: 10.1128/AAC.26.1.82.
Transposon Tn7 codes for resistance to trimethoprim and streptomycin. For detection of Tn7 by DNA-DNA hybridization, two recombinant plasmids were constructed. The former contained a 1-kilobase BamHI fragment and the latter contained a 4.3-kilobase EcoRI-BamHI fragment of Tn7. These DNA fragments, which did not include the drug resistance genes, were used as probes for detecting Tn7-like sequences in bacterial strains by colony hybridization. They hybridized strongly to bacterial DNA known to carry Tn7 but not to DNA known to carry transposons other than Tn7. These probes were used to study the occurrence of Tn7 in bacterial strains isolated in the Turku City Hospital in Finland. Transposon Tn7 was present in 47.2% of 199 trimethoprim-resistant enterobacteria (MIC greater than or equal to 8 micrograms/ml). Among the 69 Proteus mirabilis strains studied, 75% contained Tn7, although none of these strains transferred trimethoprim resistance in conjugation tests. The reliability of colony hybridization was further confirmed by Southern hybridization to detect the Tn7-specific 2.6-kilobase HindIII restriction fragment. Colony hybridization proved to be a sensitive and rapid method for detecting Tn7-determined sequences.
转座子Tn7编码对甲氧苄啶和链霉素的抗性。为了通过DNA-DNA杂交检测Tn7,构建了两种重组质粒。前者含有一个1千碱基的BamHI片段,后者含有一个4.3千碱基的Tn7的EcoRI-BamHI片段。这些不包括耐药基因的DNA片段被用作探针,通过菌落杂交检测细菌菌株中Tn7样序列。它们与已知携带Tn7的细菌DNA强烈杂交,但不与已知携带除Tn7以外的转座子的DNA杂交。这些探针用于研究芬兰图尔库市医院分离的细菌菌株中Tn7的出现情况。在199株对甲氧苄啶耐药的肠杆菌(最低抑菌浓度大于或等于8微克/毫升)中,47.2%存在转座子Tn7。在所研究的69株奇异变形杆菌菌株中,75%含有Tn7,尽管这些菌株在接合试验中均未转移甲氧苄啶抗性。通过Southern杂交检测Tn7特异性的2.6千碱基HindIII限制性片段,进一步证实了菌落杂交的可靠性。菌落杂交被证明是检测Tn7决定序列的一种灵敏且快速的方法。