Loss of protein expression following double transformation - Protein expression post transformation (Feb/12/2011 )

Hi all,

Just to fill you in on the results of my work.

I tried culturing the bacteria then half was spun down and resuspended in fresh media with ampicillin and the rest left in original media. When I induced protein expression I did that at either 37, 26, and 15 degrees Celcius. Then I ran the bacteria through a flow cytometer to assess GFP production. There was very little change resulting from the media change (some up and others down). The temperature had some impact and optimal results were obtained from the 26 degree cultures.

My options now are to find an alternate bacterial expression vector with a different origin of replication (difficult to find one) or look for a vector with dual expression in mammalian and bacterial hosts.