Konstrukcija eksperimentalnog sustava za ispitivanje mehanizma popravka dvolančanog loma DNA u bakterije Escherichia coli

master's thesis

• Title (english): Construction of an experimental system for study of the mechanism of DNA double-strand break repair in Escherichia coli
• Author: Petra Lukić
• Mentor: Zoran Zgaga (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); Zagreb
• Defense date and country: 2006-04-27, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Dvolančani lom u molekuli DNA jedno je od najopasnijih oštećenja koje se može pojaviti u staničnom genomu. Nepopravljeni dvolančani lom može dovesti do gubitka oštećenog kromosoma, a u slučaju nepreciznog popravka može doći do gubitka ili promjene dijela genetičke upute. U ovom radu su konstruirani različiti sojevi bakterije E. coli u kojima je moguće pratiti popravak dvolančanog loma induciranog specifičnom endonukleazom I-Sce I između dviju ponovljenih sekvencija. Da bi se odredila moguća uloga proteina RecA, RecF, RecN, UmuC, UmuD'C (Pol V) i Pol IV konstruirani su sojevi koji, osim eksperimentalnog sustava za indukciju dvolančanog loma, imaju i deletirane odgovarajuće gene. Rezultati ukazuju da je popravak dvolančanog loma između dviju ponovljenih sekvencija znatno manje efikasan u E. coli nego u stanicama eukariota te da je gotovo u potpunosti ovisan o produktu gena recA, a djelomično i o recF.

Abstract (english)

Double-strand break (DSB) is one of the most dangerous lesions that can be found in the cellular genome. The absence of DSB repair leads to the chromosome loss, while error-prone repair may promote loss or alterations of the genetic information. In this work different E. coli strains were constructed in order to follow repair of the DSB induced by a sequence-specific endonuclease I-Sce I between two repeated sequences. To determine the possible role of RecA, RecF, RecN, UmuC, UmuD’C (Pol V) and Pol IV proteins in this process we also performed deletions of the corresponding genes in the E. coli genome. The results obtained indicate that the DSB repair process operating between two repeated sequences is less efficient in E. coli then in eukaryotic cells. The repair is almost completely dependent on the recA gene product and much less on the recF gene.

• Keywords: homologna rekombinacija; popravak dvolančanog loma; RecA; SOS sustav
• Keywords (english): double-strand break repair; homologous recombination; RecA; SOS system
• Language: croatian
• URN:NBN: urn:nbn:hr:159:174298
• Study programme: Title: Biotechnology; specializations in: Biochemical-Microbiology; Course: Biochemical-Microbiology; Study programme type: university; Study level: graduate; Academic / professional title: diplomirani inženjer biotehnologije; smjer biokemijsko-mikrobiološki (diplomirani inženjer biotehnologije; smjer biokemijsko-mikrobiološki)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-09-08 12:40:47