Mjerenje razine oksidativnog stresa u vinske mušice pomoću ekspresije gena Hsp70 i Hsp22 obilježenih zelenim fluorescencijskim proteinom

master's thesis

• Title (english): Levels of chemical and social oxidative stress in Drosophila melanogaster measured by expression of Green Fluorescent Protein tagged to genes Hsp70 and Hsp22
• Author: Melina Butuči
• Mentor: Sergey Nuzhdin https://orcid.org/0000-0002-9963-151X (mentor)
• Mentor: Mirjana Pavlica (mentor)
• Committee member: Mirjana Pavlica (predsjednik povjerenstva)
• Committee member: Zlatko Liber (član povjerenstva)
• Committee member: Zdravko Dolenec (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2010, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Temeljno načelo teorije slobodnih radikala bazira se na pretpostavci da je unutarstanična produkcija reaktivnih intermedijera kisika (ROS) odlučujući faktor u određivanju životnog vijeka. Proteini toplinskog šoka (Hsp) također su sve više umiješani u fenotipove koji pokazuju starenje te u kontrolu životnog vijeka u raznih vrsta organizama. Zeleni fluorescencijski protein (GFP) često se koristi kao reporter ekspresije gena. U ovom istraživanju GFP je spojen s genima hsp70 i hsp22 genima, čiju ekspresiju aktivira oksidacijski stres. Moguće je pratiti/ mjeriti fluorescenciju tj. ekspresiju gena u vinskih mušica. U ovom radu mjerena je razina oksidativnog stresa u vinske mušice pomoću ekspresije gena hsp70 i hsp22 obilježenih GFP-om. Oksidacijski stres u mušica izazvan je: 1. vodikovim peroksidom i 2. Različitom organizacijom socijalnih grupa mušica s obzirom na omjer spolova. Rezultati su pokazali da postoji značajan utjecaj socijalnih grupa i kemijskog okoliša u nastanku oksidacijskog stresa, posebice u ženki vinske mušice. Općenito, vinske mušice na vodikovom peroksidu pokazale su veću ekspresiju Hspa. Međutim, nije uočena razlika u ekspresiji između različitih genotipova.

Abstract (english)

The free radical theory of aging postulates that the production of intracellular reactive oxygen species (ROS) is the major determinant of the life span. Heat- shock proteins (Hsp) are increasingly being implicated in aging phenotypes and control of life span across species as well. Green Fluorescent Protein (GFP) is frequently used as a reporter of a gene expression. Here, GFP is fused with the hsp70 and hsp22 genes that are known to be expressed during oxidative stress. It is possible to measure the fluorescence / gene expression in the fruit flies. In this work the levels of chemical and social oxidative stress in fruit flies were measured by expression of GFP tagged to the hsp70 and hsp22 genes. The flies are conditioned in two different ways: first, the oxidative stress was caused by hydrogen peroxide; second, the flies were organized in different social groups, in various ratios males to females. Results showed that there are significant effects of the social group and chemical environment in causing oxidative stress, especially in female fruit flies. In general, flies on hydrogen peroxide had higher Hsp expression. However, the actual genotype never came out as an important factor.

• Keywords: zeleni fluorescencijski protein (GFP); proteini toplinskog šoka (Hsp); reaktivni intermedijeri kisika (ROS); oksidacijski stres; socijalne grupe
• Keywords (english): GFP; Hsp; ROS; oxidative stress; social environment
• Language: croatian
• URN:NBN: urn:nbn:hr:217:699607
• Study programme: Title: Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra edukacije biologije (magistar/magistra edukacije biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2018-03-22 10:27:34