Rerun of the Test Expression of Huntingtin Domain Constructs (2016/07/06)

The last run of experiments where I have been trying to make huntingtin protein in insect cells have not been very successful. However, this has not deterred me! Looking back on the previous test expression of all of the different huntingtin fragments, there are a number of issues with the data so I decided to repeat the experiment which has taken quite a while.

The insect cell team at the SGC have repeated the work keeping a really close eye on everything and we have had success – we are now able to make small portions of the huntingtin protein in the lab! This is a really important step forward in trying to understand what the huntingtin protein looks like as we are now able to produce the protein with which to run the subsequent experiments on. However, we still need to validate whether we can scale up the making of the protein in the lab so that we are making meaningful quantities.

Below is a summary of the positive data:

TestX

The full-length huntingtin protein is shown at the bottom as the scale from 1-3144. Small fragments of the huntingtin protein starting at amino acid 80, 85 or 97 and ending at 387, 399, 419 or 428 can be expressed in the lab. We can see the protein produced when we analyse the sample on a gel – these are the blue bands highlighted by the red arrows, the fatter the blue band, the more protein that has been purified.

All of the details of these experiments including detailed methods are published on Zenodo. More to follow soon!

 

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