Cloning and test expression of HTT fragment clones - 2019/02/28

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.