Release of β-galactosidase from E. coli by a plasmid containing a temperature sensitive lytic function

• Published in: Biotechnology letters Vol. 11; no. 12; pp. 845 - 850
• Main Authors: DABORA, R. L, EBERIEL, D. T, COONEY, C. L
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 01.01.1989
• Subjects: Analytical, structural and metabolic biochemistry; beta -galactosidase; Biological and medical sciences; Biotechnology; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic technics; Hydrolases; lysis; Methods. Procedures. Technologies; plasmids; Vectors (cloning, transfer, expression). Insertion sequences and transposons; Construction; Temperature; Enzyme; Escherichia coli; β-D-Galactosidase; Plasmid; Enzymatic activity; Gene; Lysis; Bacteria; Kinetics; Escherichieae; Release; Enterobacteriaceae
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/BF01026838

We report the construction of a plasmid which carries the Phi X174 lysis gene E downstream from the lambda pL promoter. This plasmid is capable of triggering lysis in E. coli when the growth temperature is raised from 30 degree C to 42 degree C. The kinetics of release of beta -galactosidase and intracellular protein have been determined, as well as the decrease in cell viability upon induction of lysis. In addition, beta -galactosidase can be released efficiently after induction by a short sonication period. Both methods allow efficient release of beta -galactosidase.