a couple of quick things. This sounds like more of a protein-folding
issue. The lack of expression is probably caused by the cells clearing
of aggregated protein. You want to have conditions where the protein can
properly fold.
1) have you considered periplasmic expression by adding a periplas
Following on from Stephen's comment. Jena Biosciences sell a kit that
allows you to do it quickly and easily.
check it out here:
http://www.jenabioscience.com/cms/en/1/catalog/529_jbs_methylation_kit.html
We have had success in our lab using lysine methylation to crystallise
an otherwise reca
