Dear Opher,
It was also our idea how to get over it since several years ago. We
did it based on a philosophy that any bacteria infected with a phage
will not be infected by the same phage again. Therefore, we let our
BL21/DE3 strain infected in the lab and when most were died, some
cells were survived. We picked these to make the competent cells for
expression constructs. We believe these actually contained phages.
In order to avoid this bomb to blow up, we managed to get a commercial
strain (TonA and TonB deletion) purchased from Sigma USA but found it
useless. I wonder how you screened your phage-resistant strains.
In addition, it is our regular practice to use bleach to extensively
disinfect all surfaces, including benches, used flasks, instruments
and floors. Would virkon work more efficiently in our case?
Sincerely,
Aidong
On Mar 3, 2013, at 3:36 AM, Opher Gileadi wrote:
Hi,
We had a massive phage infection at the SGC in 2004; all our washing
and sterilization efforts could only solve the problem temporarily. I
then recovered a phage-resistant subclone of BL21(DE3), and prepared
derivative strains with pRARE2 (the tRNA plasmid in Rosetta2) or other
plasmids. We have expressed thousands of protein since and never had a
recurrence of phage problems. Although the resistance is to T1 phages,
it seems these are the most common lab infections.
Write me if you want the strains.
Opher
Aidong Han, Ph.D
Department of Biomedical Sciences
School of Life Sciences
Xiamen University
3 South Xiangan Road
Xiangan, Xiamen 361102
China
Phone: 0592-218-8172 (O)
0592-218-8173 (L)
Web: http://life.xmu.edu.cn/adhanlab/
