Site directed mutagenesis GFP to RFP - (Oct/22/2005 )

Hi friends, we've a vector with GFP. We r planning to change GFP into Red FP and Yellow FP using site directed Mutagenesis. Waiting for suggestions/advices. (sequences,tips,clues).Thanx

The sequences of many fluorescent proteins is found on www.clontech.com. I'm not sure if it's legal to just do it all on your own (making the exact same genes), it might be.

Are you experienced with Site directed mutagenesis? If not, you can check for instance at www.stratagene.com for tips.

Am not very fluent in site directed mutagenesis but hope the pages suggeste dby u will help initialyy,thank u.

The sequences of the red fluorescent proteins and GFP arecompletely different. The yellow and cyan proteins are mutated forms of GFP, however. I believe one or two mutations could change the color of the GFP gene, but would not create the "standard" YFP or CFP. Nonetheless, this might be adequate for your needs. See for example this site:

http://www.microscopyu.com/articles/livece...ng/fpintro.html

I found it with the keywords "venus topaz gfp" and google, which finds several other sites as well.

Why do the hassle of mutagenesis and then cloning.

Clontech sells multiple RFP plasmids and a guy in Califormia has developed a rainbow of colours now or soon to be available from BD Clontech.

As Varius said, there may be some patenting issues here.