Characterization of Escherichia coli lysis using a family of chimeric E-L genes.
Gene E-L, a chimeric lysis construct from bacteriophages phi X174 and MS2 lysis proteins E and L, respectively, was subjected to internal deletions to create a series of new E-L clones with altered lysis or killing properties. The lytic activities of the parental genes E. L. E-L and the internal truncated forms of E-L were investigated in this study to characterize the different lysis mechanisms, based on differences in the architecture of the different membrane spanning domains. Electron microscopy and release of marker enzymes for the cytoplasmic and periplasmic spaces revealed that two different lysis mechanisms can be distinguished depending on penetrating of the proteins either the inner membrane or the inner and outer membranes of Escherichia coli. Several candidates, which share efficient lysis properties, have biotechnological applications in terms of cell disruption.
Institute of Microbiology and Genetics, University of Vienna, Austria. emily@gem.univie.ac.at
A
A
Witte
G R
GR
Reisinger
W
W
Säckl
G
G
Wanner
W
W
Lubitz
D001433:Bacteriolysis / Q000502:physiology*; D001483:Base Sequence; D004926:Escherichia coli / Q000648:ultrastructure / Q000821:virology*; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000502:physiology*; D013997:Time Factors; D014170:Transformation, Genetic; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology*; D001616:beta-Galactosidase / Q000032:analysis; D001618:beta-Lactamases / Q000032:analysis
C029555:E protein, bacteriophage X174; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C037227:lysis protein, Enterobacterio phage MS2; D001616:beta-Galactosidase; D001618:beta-Lactamases