An in vitro study of the expression, purification and intramolecular interactions of Hepatoma Up Regulated Protein (HURP) [Greek]

By Eva Ka

Bachelor Thesis from the year 2011 in the subject Biology - Micro- and Molecular Biology, grade: A+ or 10.0, Democritus University Of Thrace (Molecular Biology and Genetics), language: Greek, Modern (1453-), abstract: This manuscript is a diploma undergraduate thesis written in Greek; Research was conducted at the University of Thrace, at the Department of Molecular Biology and Genetics. Hepatoma Up Regulated Protein (HURP) is a microtubule-associated protein essential for the assembly of the mitotic spindle. HURP binds microtubules via its N-terminal domain, which is inhibited by HURP's C-terminal tail. The kinase Aurora A activates HURP by phosphorylating its C-terminus, which dissociates from the N-terminal microtubule binding domain. This diploma thesis explores how the autoinhibitory interaction between the N- and C-termini of HURP affect binding to proteins with key functions in spindle assembly (e.g., the mitotic motor Eg5 and the microtubule associated proteins TPX2 andXMAP125). I used molecular cloning techniques to generate recombinant GST- and His tagged chimeras of HURP's N- and C-termini. I optimized conditions for the expression of these proteins in bacteria and their purification by liquid chromatography methods using an FPLC system. I performed pull-down assays by incubating the recombinant proteins with HeLa cell extracts and ran western blots to examine the interactions of HURP with Eg5, TPX2 and XMAP125.

• Format: Paperback
• Language: Greek
• ISBN: 3668190437
• ISBN13: 9783668190436
• Release Date: April 2016
• Publisher: Grin Verlag
• Length: 108 Pages
• Weight: 0.33 lbs.
• Dimensions: 0.3" x 5.8" x 8.3"