Recently I have been busy making lots of huntingtin protein samples for various experiments I have planned shortly to work out what the protein is doing with respect to binding DNA and other nucleic acid material.
I have been using constructs we made in house which have either N-terminal or C-terminal FLAG tags and a range of different polyQ lengths.
A) pBacMam vector map B) 28 HTT expression constructs with different polyQ lengths were generated with either N or C terminal FLAG-tags, C) FLAG-tags are appended to either end of the HTT expression construct with minimal additional sequence.
With these I have expressed full-length huntingtin protein in mammalian cells here, here and here, as well as making protein in insect cells. I have also been working on optimising huntingtin exon 1 protein expression which at first, proved challenging, but in the end I got some nice samples.
More coming soon!