Today’s post details work which follows on from the test expression experiments where we screened different sections of the huntingtin gene to see which successfully permitted soluble expression of protein in our insect cell expression system. The aim of these experiments is to try and see whether we can make parts of the huntingtin protein recombinantly (in the test tube so to speak). If we can make these chunks of the huntingtin protein we will then begin to characterise them to understand further how they are folded, what they might be doing in the cell and also what their structure i.e. what do they look like? One step at a time though…!
Four constructs which were positive hits from the test expression experiment, were scaled up to see if we could, firstly, reproduce the test expression data and secondly, to see if we could make reasonable quantities of these proteins of good quality.
You can read all about this work on Zenodo. Please feel free to let me know what you think. There is still a way to go before we have well behaving samples.
To complete these experiments, I was helped by Mani Ravichandran and Chelsea Cheng, pictured above. Thanks to both of them for their meticulous and hard work.