Ispitivanje interakcija halogeniranih i nehalogeniranih derivata cijanina s dvolančanim DNA i RNA spektroskopskim metodama

master's thesis

• Title (english): Study of interactions of halogenated and non-halogenated cyanine derivatives with double-stranded DNA and RNA by spectrosopic methods
• Author: Petra Mihovilović
• Mentor: Marijana Radić Stojković (mentor)
• Committee member: Iva Juranović Cindrić (predsjednik povjerenstva)
• Committee member: Ines Primožič (član povjerenstva)
• Committee member: Marko Močibob (član povjerenstva)
• Committee member: Tomislav Jednačak (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Chemistry); Zagreb
• Defense date and country: 2021-10-22, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Chemistry

Abstract

Cijaninske boje, kao poznate fluorescentne probe, pronalaze primjenu u specifičnoj detekciji
nukleinskih kiselina. U okviru ovog diplomskog rada spektroskopski su okarakterizirane tri
novosintetizirane cijaninske boje. Otopine spojeva posjeduju karakteristični UV/Vis spektar u
valnom području od 220 nm do 600 nm. Intrinzično nefluorescentni cijanini, odlikuju se izrazitim
fluorimetrijskim odgovorom uslijed vezanja na polinukleotide. Interakcije cijaninskih boja s
dvolančanim polinukleotidima ispitane su standardnim spektrofotometrijskim metodama.
Eksperimentima temperaturnog mekšanja i titracijama metodom cirkularnog dikroizma utvrđen
je način vezanja spojeva na polinukleotide. Obradom fluorimetrijskih titracija pomoću
Scatchardove jednadžbe metodom nelinearne regresije dobivene su konstante stabilnosti
kompleksa cijaninskih boja s polinukleotidima. Cijanini su pokazali selektivnost u odnosu na na
sekvencu i sekundarnu strukturu nukleinskih kiselina. Dobiveni rezultati ukazuju na mješovit
način vezanja cijaninskih boja, u kojemu uz interkalaciju dolazi do vezanja dimera ili većih
agregata u utore polinukleotida

Abstract (english)

Cyanine dyes, as known fluorescent probes, find application in the specific detection of nucleic
acids. Three newly synthesized cyanine dyes were characterized spectroscopically as part of
this thesis. Aqueous solutions of compounds absorbed in the UV/Vis region 220-600 nm.
Intrinsically non-fluorescent cyanines are characterized by a pronounced fluorimetric response
upon binding to polynucleotides. Interactions of cyanine dyes with double-stranded
polynucleotides were examined by standard spectrophotometric methods. The mode of binding
of compounds to polynucleotides was determined by temperature melting experiments and CD
titrations. Stability constants of the complexes with polynucleotides were obtained by processing
fluorimetric titrations using the Scatchard equation. Cyanines have shown selectivity with
respect to the sequence and secondary structure of nucleic acids. The obtained results also
indicate a coexistence of several binding modes, in which intercalation is accompanied by
binding of dimer or higher-order aggregates inside the polynucleotide grooves.

• Keywords: apsorbancija; cirkularni dikroizam; cijaninske boje; DNA; fluorescentne probe; RNA
• Keywords (english): absorbance; circular dichroism; cyanine dyes; DNA; fluorescent probes; RNA
• Language: croatian
• URN:NBN: urn:nbn:hr:217:889313
• Project: Number: IP-2018-01-4694; Title: Molekularno prepoznavanje DNA:RNA hibridnih i višelančanih struktura u bioanalitičkim i in vitro sustavima; Acronym: DNARNAHyB-MolBio; Leader: Marijana Radić Stojković; Jurisdiction: Croatia; Funder: HRZZ; Funding stream: IP
• Study programme: Title: Graduate university studies in chemistry; specializations in: Research programme; Course: Research programme; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra kemije (magistar/magistra kemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2023-10-22
• Created on: 2021-11-08 11:25:57