I would like to perform a SILAC experiment in E.coli. does anybody have a good method for it?
thank you in advance,
I have asked a colleague from the MS group and post his answer below. I hope there is something useful in the links that he gave me. I will forward the email to your address, so you cvan get the attached pdf too.
"I do not have a protocol for SILAC in E. coli specifically as I never used SILAC in E. coli myself but have some experience labelling E. coli with artificial amino acids.
Depending on the application, proteomics or recombinant protein production, I guess the label could differ according to the question. Although for quantitative proteomics Lys and Arg as a combination with Trypsin for digestion are most commonly used.
I attached a pdf from a protocol paper. And also the links below to some papers in which E. coli is used can be of interest.
For the rest: Any minimal medium should work. The question is whether or not to use an auxotrophic or a prototroph and supress endogenous synthesis by adding the amino acids to the medium.
For proteomics the conversion of arginine to proline is a potential pitfall with several solutions, see links below:
Hope this enough info to start with,
Please tell your colleague to feel free to contact me if he has follow up questions."