The use of agricultural biomass-based fertilizers, and the release of feces into the environment leads to last-lasting pollution of antibiotic resistance genes that cannot be removed from waters via traditional methods, resulting in significant health threats. To solve this issue, an antibiotic resistance gene removal method was proposed and tested that used sequence-specific DNA-binding designer zinc finger proteins, which target an 18-bp DNA sequence for specific antibiotic resistance gene binding and removal. Targeting the sulfonamide-resistant sul1 gene, sul1-binding zinc-finger protein was designed, overexpressed, and purified. This protein showed specific binding with sul1 over tetA that do not have the targeted sequence. This protein was further immobilized on agarose-based resins to prepare a sul1-removal column. When loaded with 10 mg protein, this column can remove over 99 % sul1 in water, suggesting high efficiency. This work presents a new method attempting to eliminate environmental and health threats posed by antibiotic resistance genes.

中文翻译：

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使用设计锌指蛋白高效去除抗生素耐药基因


使用基于农业生物质的肥料以及将粪便释放到环境中会导致抗生素耐药基因的持久污染，这些基因无法通过传统方法从水中去除，从而导致严重的健康威胁。为了解决这个问题，提出了一种抗生素耐药基因去除方法并进行了测试，该方法使用序列特异性 DNA 结合设计锌指蛋白，该方法靶向 18 bp DNA 序列进行特异性抗生素耐药基因结合和去除。靶向磺胺类抗性 sul1 基因，设计、过表达和纯化 sul1 结合锌指蛋白。该蛋白显示出与没有靶向序列的 tetA 上的 sul1 的特异性结合。将这种蛋白质进一步固定在基于琼脂糖的树脂上，以制备 sul1 去除柱。当加载 10 mg 蛋白质时，该色谱柱可以去除超过 99% 的水中 sul1，表明效率很高。这项工作提出了一种新方法，试图消除抗生素耐药基因带来的环境和健康威胁。