This past week I have been working hard to make lots of protein for future experiments.
- I have successfully purified full-length C-terminally tagged huntingtin. Having the FLAG tag on the C-terminal end of the protein seems to reduce the aggregation of the sample – I have now seen this consistently in 2 different purifications. I plan to optimize EM sample preparation with this sample.
- Purification of huntingtin domain constructs remains challenging but I did manage to purify some very clean huntingtin protein spanning P80-G428. I hope to repeat this purification and set up some crystal trays in the coming weeks.
- HMGB1 remains a challenging protein to purify and I have yet to optimize this purification procedure but I will repeat this experiment soon. Third time lucky?
- Three different p53 constructs were expressed and purified, now I need to test if p53 is a direct interacter of huntingtin.
Now I have lots of protein in hand, I can do some more exciting functional and structural experiments. Stay tuned to see what I find out.
Great job!
I am a postdoc in UTSW, and I plan to work on Huntington’s disease.
Could you give me some plasmid?
pBacMam-diex-lic-huntingtin with a C-terminal tag instead.
pFastBac-Huntingtin and pFastBac-mutant huntingtin.
Thanks,
Xiao
Absolutely! I will drop you an email this coming week to arrange. Thanks for your interest.