Transformation in Staphylococcus epidermidis - (Jan/08/2015 )

Dear all,

              i am a graduate student from India. i have been trying to transform a Staphylococcus epidermidis specific plasmid pCM29 into the S. epidermidis CVD085C strain, i am working with through electroporation (Augustin and Gotz, 1990). i could not find any other protocols apart from electroporation. But, i could not transform the plasmid into the strain through electropration. Can any body suggest how i could approach with this problem? is there any efficient method for transformation of pCM29 in to S. epidermidis  ? Kindly help.

Some possible problems include restriction enzymes present in your target cell. If you have sequence, you can identify the possible enzymes. Check the Rebase database, http://rebase.neb.com

You may be lucky in having available methylases for these enzymes (more likely not).

You may be able to engineer the cut sites out of your vector. Transformation with RE sites will still work, but often at dramatically lower efficiency.

You could try some alternative transformation techniques. Transformation by conjugation, using pir+ coli cells (S17-1 strain) is often successful for difficult transformation.