10.5281/zenodo.2600051
https://zenodo.org/records/2600051
oai:zenodo.org:2600051
Harding, Rachel
Rachel
Harding
0000-0002-1134-391X
Structural Genomics Consortium, University of Toronto
Hutchinson, Ashley
Ashley
Hutchinson
Structural Genomics Consortium, University of Toronto
Seitova, Alma
Alma
Seitova
Structural Genomics Consortium, University of Toronto
Arrowsmith, Cheryl
Cheryl
Arrowsmith
Structural Genomics Consortium, University of Toronto
Lee, Matt
Matt
Lee
CHDI Foundation
Cloning and test expression of HTT fragment clones - 2019/02/28
Zenodo
2019
Huntingtin
Protein expression
Protein purification
X-ray crystallography
2019-02-28
10.5281/zenodo.2600050
https://zenodo.org/communities/labscribbles
https://zenodo.org/communities/sgc-opennotebook
https://zenodo.org/communities/openlabnotebooks
Creative Commons Attribution 4.0 International
Project: High resolution structural analysis of purified HTT proteins
Experiment: Cloning and test expression of HTT fragment clones
Date completed: 2019/02/28
Rationale: I aim to complement our previous work on purifying stable regions of huntingtin by cloning further regions of huntingtin corresponding to discrete HEAT repeats for eukaryotic expression. Constructs will initially be screened by BVES in insect sf9 cells. High quality constructs will undergo extensive crystallization experiments and structure determination by X-ray crystallography using well established high-throughput and systematic protocols in place at the SGC. Any solved structures may allow generation of a pseudoatomic resolution (<4 Å) model of the HTT protein using the cryoEM model as a guide.
Dr. Harding is the recipient of the Huntington's Disease Society of America Berman Topper Career Development Fellowship which funds and supports this research, in addition to generous funding from the CHDI Foundation and the Huntington Society of Canada. The SGC is a registered charity (number 1097737) that receives funds from AbbVie, Bayer Pharma AG, Boehringer Ingelheim, Canada Foundation for Innovation, Eshelman Institute for Innovation, Genome Canada through Ontario Genomics Institute [OGI-055], Innovative Medicines Initiative (EU/EFPIA) [ULTRA-DD grant no. 115766], Janssen, Merck KGaA, Darmstadt, Germany, MSD, Novartis Pharma AG, Ontario Ministry of Research, Innovation and Science (MRIS), Pfizer, São Paulo Research Foundation-FAPESP, Takeda, and Wellcome.