A freeze-thaw method for disintegration of Escherichia coli cells producing T7 lysozyme used in pBAD expression systems.
The pLysN plasmid containing the T7 lysozyme gene under control of the lac promoter was constructed to facilitate cell disintegration after expression of recombinant proteins in arabinose-induced expression systems. The usefulness of this plasmid was tested in Escherichia coli TOP10 and E. coli LMG194 cells carrying pBADMHADgeSSB plasmid containing Deinococcus geothermalis SSB protein gene under control of the araBAD promoter. The results showed that low-level expression of T7 lysozyme did not interfere with the target SSB protein production, and that the freezing-thawing treatment was sufficient for disruption of the E. coli cells producing low amounts of T7 lysozyme.
Wanarska
Marta
M
Department of Microbiology, Chemical Faculty, Gdańsk University of Technology, Gdańsk, Poland. martka.chem@wp.pl
Hildebrandt
Piotr
P
Kur
Józef
J
D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D034301:Deinococcus / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005615:Freezing; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism*
D004268:DNA-Binding Proteins; D009238:N-Acetylmuramoyl-L-alanine Amidase