Success with T7

During the past few days I have conducted an experiment to test whether cooling T7 RNA polymerase to 25º allows sxy gene transcripts to fold in vitro as they do in vivo. Although I have not directly tested whether sxy transcripts are folding into the structures we have previously determined experimentally, conducting in vitro transcription/translation at 25º yields results much more in line with what we have detected in vivo. The new results are that sxy-1 is translated at high levels, whereas wildtype and sxy-7 transcripts are not. I am currently repeating the experiment and am also allowing the reactions to proceed for up to 4 hrs to see whether a difference emerges between the two poorly-translated transcripts (WT and sxy-7).