The Use of Analytical Isoelectric Focusing for Detection and Identification of -Lactamases

作者：Margaret Matthew, ANNE M. HARRIS, Matthew J. Marshall, Gillian Ross · 发表于：Journal of General Microbiology · 年份：1975 · DOI：10.1099/00221287-88-1-169 · 被引用次数：691 · 研究领域：Genomics and Phylogenetic Studies、Microfluidic and Capillary Electrophoresis Applications、Biosensors and Analytical Detection

BETA-Lactamases (EC. 3.5.2.6) from strains of Gram-negative bacteria have been studied using analytical isoelectric focusing. This permits a visual comparison of the patterns of beta-lactamase bands produced by enzymes from different organisms. Purification of crude intracellular preparations is unnecessary and the technique is sufficiently sensitive to demonstrate beta-lactamase in mutants previously reported to lack the enzyme. R that have not been distinguished from one another biochemically or immunologically can be differentiated by isoelectric focusing. Conversely, the enzymes specified by the R factors RTEM, R1 and RGN14, with identical isoelectric focusing patterns have the same biochemical properties. Chromosomal and R-factor-mediated beta-lactamases from single strains have been separated and their identities confirmed by immunoisoelectric focusing. R factor-mediated enzymes gave identical isoelectric focusing patterns irrespective of the host strain. Isoelectric focusing can therefore be used to observe the transfer of beta-lactamases carried by R factors.