Bacillus stearothermophilus transformation

rafael counago ralemico at hotmail.com
Thu Feb 8 16:48:58 EST 2001


Hi,

I know there are some articles in this mail list about this issue, but ...

I am trying to perform homologous recombination on B. stearothermophilus. My 
major problem is that I cannot transform the bugs! I tried the protoplast 
protocol (Dr. Welker) with the NUB3621 strain, but did not have any success. 
Even when using pUB110 as a positive control and different lysozyme 
concentrations. I've been keeping the cells at high temperatures and 
centrifuging at room temperature.

Does anybody out there have any ideas/advise on how to make this protocol to 
work?

I am also about to give the electroporation protocol a try. Are there 
anybody that have been working on that and can give some advise?

I very much appreciate any help.

Thanks,

Rafael
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