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An in vitro study of the expression, purification and intramolecular interactions of Hepatoma Up Regulated Protein (HURP)
Buch (Taschenbuch, Griechisch)
Fr.55.90
inkl. gesetzl. MwSt.Beschreibung
Details
Einband
Taschenbuch
Erscheinungsdatum
19.04.2016
Verlag
GRINSeitenzahl
108
Maße (L/B/H)
21/14.8/0.8 cm
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: Modern Greek, 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.