用于氨基糖苷类修饰酶的微孔板磷酸纤维素结合测定法。
Microplate phosphocellulose binding assay for aminoglycoside-modifying enzymes.
作者信息
Cooksey R C, Metchock B G, Thornsberry C
出版信息
Antimicrob Agents Chemother. 1986 Dec;30(6):883-7. doi: 10.1128/AAC.30.6.883.
Abstract
We modified the phosphocellulose binding assay for aminoglycoside-modifying enzymes (AMEs) by use of microdilution plates and a multichannel micropipette. Batteries of aminoglycoside substrates for screening organisms for the presence of AMEs as well as for subclassifying enzymes were prepared and stored in microdilution plates. When tested in parallel with the conventional tube reaction assay, the microplate assay yielded comparable radioactive counts and therefore equally correct identifications of AMEs in 32 isolates representing nine bacterial species. Other modifications, such as multichannel dispensing of crude enzyme preparations and radioisotopic precursors, provided a more rapid, convenient, and less expensive means of examining large collections of organisms for AMEs.
摘要
我们通过使用微量稀释板和多通道微量移液器改进了用于氨基糖苷修饰酶（AMEs）的磷酸纤维素结合测定法。制备了用于筛选生物体中AMEs以及对酶进行亚分类的一系列氨基糖苷底物，并储存在微量稀释板中。当与传统的试管反应测定法平行测试时，微孔板测定法产生了相当的放射性计数，因此在代表9种细菌的32株分离物中对AMEs的鉴定同样正确。其他改进，如粗酶制剂和放射性同位素前体的多通道分配，为检测大量生物体中的AMEs提供了一种更快速、方便且成本更低的方法。
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