Konstrukcija i provjera plazmida pAB9-126S8

undergraduate thesis

• Title (english): Construction and verification of the plasmid pAB9-126S8
• Author: Fran Andrašec
• Mentor: Ivan-Krešimir Svetec (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Biology and Microbial Genetics); Zagreb
• Defense date and country: 2012-07-25, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Cilj ovog rada bio je tehnikama genetičkog inženjerstva konstruirati plazmid pAB9-126S8 koji sadrži dva obrnuta ponavljanja od kojih je svako dugo 63 pb s razmaknicom od 8 pb. Plazmid je konstruiran modifikacijom plazmida pAB9-114 koji sadrži obrnuta ponavljanja duljine 57 pb bez razmaknice (palindrom). Palindrom je modificiran na način da je u njegov centar umetnut fragment čime su ruke palindroma produžene s 57 na 63 pb te je uvedena razmaknica. Nakon transformacije bakterije Escherichia coli u kojoj se plazmid može održavati i umnažati, provedena je analiza koja je omogućila razlikovanje plazmida prema broju umetnutih inserata kako bi se odabrali transformanti koji nose željeni plazmid s jednim insertom. Od deset analiziranih transformanata, devet ih je sadržavalo plazmid s jednom kopijom inserta.

Abstract (english)

The goal of this work was to construct the plasmid pAB9-126S8 which contains two inverted repeats of 63 bp with a spacer of 8 bp using the methods of genetic engineering. This was achieved by modification of an existing plasmid pAB9-114 which contains two inverted repeats, each 57 bp long without a spacer (palindrome). The palindrome was modified by inserting a fragment which extended the hands of the palindrome from 57 to 63 bp and introduced a spacer. Following the transformation of the host Escherichia coli in which the plasmid can be maintained and replicated, an analysis was conducted which made it possible to distinguish plasmids based on the number of inserted fragments they contain in order to select the transformants which carry the desired plasmid with a single insert. Of the ten transformants which were analysed, nine carried a plasmid with a single copy of the insert.

• Keywords: Escherichia coli; plazmid; palindrom; razmaknica; transformacija
• Keywords (english): Escherichia coli; plasmid; palindrome; spacer; transformation
• Language: croatian
• URN:NBN: urn:nbn:hr:159:821798
• Study programme: Title: Biotechnology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-05-16 13:51:18