broad spectrum transposon for gram positive bacteria - (Nov/29/2005 )

Hi,

i have been trying for months to mutagenize/transform a bacteria I isolated from soil and latter found to be corynebacteruim variabilis. does someone know how to prepare electrocompetent corynebacterium variablis cells? is there a universal transposon for gram positive bacteria?
thanks for you attention

You should look at the Epicentre transposome system. It creates DNA fragments complexed with transposase protein which can be directly electroporated into cells, avoiding issues of expression of the transposase in your organism. There are standard versions with kan and Trimethoprim resistance available, and easy mechanisms for constructing your own insertion fragments. http://www.epibio.com