A freeze-thaw method for disintegration of Escherichia coli cells producing T7 lysozyme used in pBAD expression systems

• Authors: Marta Wanarska , Piotr Hildebrandt , Józef Kur
• Languages of publication: EN

Abstracts

EN

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.

Keywords

EN

pBAD expression systems; disintegration; T7 lysozyme

• Journal: Acta Biochimica Polonica
• Year: 2007
• Volume: 54
• Issue: 3
• Pages: 671-672

Dates

published

2007

received

2007-08-08

revised

2007-09-10

accepted

2007-09-14

(unknown)

2007-09-17

Contributors

author

Marta Wanarska

• Department of Microbiology, Chemical Faculty, Gdańsk University of Technology, Gdańsk, Poland

author

Piotr Hildebrandt

• Department of Microbiology, Chemical Faculty, Gdańsk University of Technology, Gdańsk, Poland

author

Józef Kur

• Department of Microbiology, Chemical Faculty, Gdańsk University of Technology, Gdańsk, Poland

References

• Bollag DM, Rozycki MD, Edelstein SJ (1996) In Protein Methods, pp 27-43. Wiley-Liss Inc., New York.
• Filipkowski P, Duraj-Thatte A, Kur J (2006) Arch Microbiol 186: 129-137.
• Studier FW (1991) J Mol Biol 219: 37-44.