Kurittu J, Karp M, Korpela M
Department of Biotechnology, University of Turku, Tykistökatu 6A, FIN-20520 Turku, Finland.
Luminescence. 2000 Sep-Oct;15(5):291-7. doi: 10.1002/1522-7243(200009/10)15:5<291::AID-BIO596>3.0.CO;2-B.
The performance of two bioluminescent Escherichia coli K-12 strains for the specific detection of the tetracycline family of antimicrobial agents was compared, and the analytical applicability of one of the strains was preliminarily evaluated. One sensor plasmid contained the bacterial luciferase operon of Photorhabdus luminescens under the control of the tetracycline-responsive element from transposon Tn10 (15). An analogous plasmid construction with firefly (Photinus pyralis) luciferase reporter gene was constructed, and these two divergent tetracycline-inducible light-emitting systems were compared for their suitability for the qualitative detection of tetracyclines. Both sensor strains behaved in a similar manner kinetically, and the most sensitive tetracycline response for both sensor strains was achieved in 90-120 min by performing the assay at 37 degrees C. The sensor strain containing the bacterial luciferase operon responded slightly more sensitively to different tetracyclines than the strain containing firefly luciferase gene. The sensor bacteria retained their inducibility in lyophilization, and freeze-dried cells detected tetracyclines as sensitively as freshly cultivated sensor cells. The preliminary results from the analysis of tetracycline-spiked pork serum samples indicated that these sensor bacteria could be used to screen veterinary samples for tetracycline residues in real-time.
比较了两种生物发光大肠杆菌K-12菌株对四环素类抗菌剂进行特异性检测的性能，并初步评估了其中一种菌株的分析适用性。一种传感器质粒含有发光杆菌的细菌荧光素酶操纵子，该操纵子受转座子Tn10的四环素响应元件控制（15）。构建了一种带有萤火虫（Photinus pyralis）荧光素酶报告基因的类似质粒结构，并比较了这两种不同的四环素诱导发光系统对四环素定性检测的适用性。两种传感器菌株在动力学上表现相似，通过在37℃下进行测定，两种传感器菌株在90-120分钟内都能获得最敏感的四环素响应。含有细菌荧光素酶操纵子的传感器菌株对不同四环素的反应比含有萤火虫荧光素酶基因的菌株稍敏感。传感器细菌在冻干过程中保持其诱导性，冻干细胞检测四环素的灵敏度与新鲜培养的传感器细胞相同。对添加四环素的猪肉血清样品的分析初步结果表明，这些传感器细菌可用于实时筛选兽药样品中的四环素残留。
