A microtiter plate-based system for the semiautomated growth and assay of bacterial cells for beta-galactosidase activity.
The introduction of automated pipetting devices, microtiter readers, and microcomputers makes it possible to significantly increase the number of enzyme assays which can be performed as part of the analysis of a biological process. A number of difficulties must be overcome in any such integrated approach based on the microtiter plate. Among these are cell lysis, temperature control, the conversion of microtiter reader optical density values to standard 1-cm path length values, and data management. The utility of such a scheme can be extended to gene regulation and bacterial genetics studies, if bacterial cell culture techniques can be incorporated into the scheme. This paper addresses these issues in the application of a semiautomated system to the study of the induction of the gyrA promoter by treatment (of a gyrA-lac operon fusion-containing strain) with a gyrase inhibitor. This system is specific to the requirements of our studies into the modulation of gene expression by DNA relaxation. The general approach, however, can be readily adapted to other studies.
Menzel
R
R
E.I. du Pont de Nemours, Central Research and Development Department, Wilmington, Delaware 19880-0328.
D001419:Bacteria / Q000201:enzymology*; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D003199:Computer Systems; D005696:Galactosidases / Q000378:metabolism*; D005786:Gene Expression Regulation; D008832:Microchemistry / Q000295:instrumentation; D010957:Plasmids; D012984:Software; D001616:beta-Galactosidase / Q000378:metabolism*
D005696:Galactosidases; D001616:beta-Galactosidase