Bacillus stearothermophilus transformation

d.studholme at ic.ac.uk d.studholme at ic.ac.uk
Wed Nov 29 14:16:34 EST 1995


Hello,

I am attempting to express a protein and also
cause gene-disruption by homologous recombination in Bacillus
stearothermophilus.

For both these tasks I need a reasonably efficient system for transformation
with plasmid DNA.

I would be extremely interested to hear from anybody working with
thermophiles who could comment on host-vector systems.

I am aware of the protoplasting/polyethylene glycol method for B. stearo
strains CU21, NUB3621 but have had very dissapointing results.
I am also aware of the electroporatable strain K1041 (Narumi et al,1994)
but again this has not worked well in my hands!

Any advice at all would be much appreciated!!!

Cheers, Dave.



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