Mohn G R
Arch Toxicol. 1984 Oct;55(4):268-71. doi: 10.1007/BF00341024.
The DNA repair host-mediated assay, in which repairable DNA damage is determined in E. coli cells present in various organs of mice exposed to genotoxic agents, was further developed to broaden the range of organs under study and to simplify the procedure of assessing differential bacterial cell survival. A pair of derivatives of E. coli K-12 strain 343/113 was constructed which differed vastly in DNA repair capacity (uvr+/rec+ vs uvrB/recA), as a means of assessing DNA damaging effects; furthermore, the strains differed in their ability to ferment lactose (delta Lac vs Lac+), so that the individual survival of both strains could be determined on a single agar medium (containing neutral red as pH indicator), on which the strains had different colony colour morphology (red, Lac+ vs white, Lac- colonies). Finally, the strains were made streptomycin-dependent, to prevent uncontrolled growth of the bacterial cells within the various organs and also to inhibit contamination of the survival agar medium by representatives of the normal intestinal microflora. The experimental procedure consisted of injecting mixtures of stationary cells of the two strains (ca. 3-5 X 10(8) viable cells per mouse) both intravenously and orally into mice, either pretreated or subsequently treated with test chemicals. Ninety minutes after injection of the bacteria, the liver, spleen, lungs, kidneys, stomach, intestine, colon, and ca. 50 microliter blood, were removed, suspended in buffer, homogenized, and the survival of the two strains determined on neutral red agar supplemented with streptomycin.(ABSTRACT TRUNCATED AT 250 WORDS)
DNA修复宿主介导试验是通过检测暴露于遗传毒性剂的小鼠各器官中存在的大肠杆菌细胞内可修复的DNA损伤来进行的。该试验得到了进一步发展,以扩大所研究器官的范围,并简化评估细菌细胞差异存活的程序。构建了一对大肠杆菌K-12菌株343/113的衍生物,它们在DNA修复能力上有很大差异(uvr+/rec+与uvrB/recA),以此作为评估DNA损伤效应的手段;此外,这两种菌株在发酵乳糖的能力上也有所不同(ΔLac与Lac+),这样就可以在单一琼脂培养基(含有中性红作为pH指示剂)上确定两种菌株的个体存活率,在该培养基上两种菌株具有不同的菌落颜色形态(红色,Lac+菌落与白色,Lac-菌落)。最后,使这些菌株依赖链霉素,以防止细菌细胞在各器官内不受控制地生长,同时也抑制正常肠道微生物群代表对存活琼脂培养基的污染。实验程序包括将两种菌株的静止期细胞混合物(每只小鼠约3 - 5×10⁸个活细胞)通过静脉和口服方式注射到经预处理或随后用测试化学品处理的小鼠体内。注射细菌90分钟后,取出肝脏、脾脏、肺、肾脏、胃、小肠、结肠和约50微升血液,悬浮于缓冲液中,匀浆,并在补充有链霉素的中性红琼脂上测定两种菌株的存活率。(摘要截断于250字)