I'm trying to electroporate a linear DNA fragment into
Rhodococcus rhodochrous. It is a gram positive soil bacteria.
The DNA fragment have 2 small fragments of Rhodococcus DNA
(around 2-4 kb) on both ends of an antibiotic resistant gene
of a plasmid. I'm trying to electroporate it into the cells
so that it can knock out a gene by homologous recombination
and insertion into the middle of that gene to disrupt it.
However, there's nothing in the literature about this that
I'm aware of. If you have any experience concerning
electroporation of this bug (like setting the parameters
to maximize the transformation) or about homologous recombination
please write me. I'm desperate to make this thing work out
because I tried it so many times and not so successful. PLEASE
HELP........Thank you very much.
*** My e-mail adress is
etn06448 at dna.bchs.uh.edubiolea at jetson.uh.edu