Molekularna diferencijacija sojeva Lactobacillus plantarum iz hrvatskih autohtonih fermentiranih proizvoda temeljena na numeričkoj analizi makrorestrikcijskih i proteinskih profila

master's thesis

• Title (english): Molecular differentiation of Lactobacillus plantarum strains from Croatian autochthonous fermented products based on numerical analysis of macrorestriction and whole cell protein patterns
• Author: Ivan Varžić
• Mentor: Višnja Bačun-Družina (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: 2012-09-13, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Proučavanje bioraznolikosti kao i diferencijacija na razini soja su važne za razumijevanje molekularne evolucije kao i za selekciju sojeva s industrijski značajnim svojstvima. Cilj ovoga rada bila je diferencijacija osam autohtonih sojeva Lactobacillus plantarum pomoću dvije različite molekularne metode kako bi utvrdili u kojoj se mjeri razlikuju sojevi iste vrste koji potječu iz različitih okolišnih niša. Denaturirajuća poliakrilamidna gel elektroforeza ukupnih staničnih proteina (WCP/SDS-PAGE) i analiza restrikcijskih fragmenata DNA elektroforezom u pulsirajućem polju (PFGE) su korištene za diferencijaciju sojeva i procjenu fenotipske i genotipske raznolikosti i varijabilnosti. Proteinski i makrorestrikcijski profili dobiveni svakom od navedenih metoda prevedeni su u dvodimenzionalnu binarnu matricu te su nakon numeričke analize podataka primjenom računalnog programa Phoretix 1D dobiveni dendrogrami koji pokazuju filogenetski odnos i relativnu sličnost između ispitivanih sojeva. Također je ispitana osjetljivost ovih sojeva na različite antibiotike. Svi sojevi su rezistentni na nalidiksinsku kiselinu, te su u većoj ili manjoj mjeri osjetljivi na tetraciklin, ampicilin, streptomicin i kloramfenikol. Rezultati su jasno ukazali na postojanje značajne raznolikosti autohtonih sojeva L. plantarum koja je vidljiva kroz njihove molekularne karakteristike kao genomski i proteinski polimorfizam.

Abstract (english)

The study of bio-diversity as well as differentiation at the strain level are important for understanding of the molecular evolution and for selection of strains with industrially relevant properties. The aim of this study was to differentiate eight autochthonous Lactobacillus plantarum strains by two different molecular methods in order to determine the extent of intraspecific differences among strains which originate from different environmental niches. Denaturing polyacrilamide gel electrophoresis of whole cell proteins (WCP/SDS-PAGE) and DNA restriction fragment analysis by pulsed-field gel electrophoresis (PFGE) were used for strain differentiation and the assessment of phenotypic and genotypic diversity and variability. Protein and macrorestriction patterns obtained from each method were converted to two-dimensional binary matrix and after numerical analysis of data by using Phoretix 1D software, dendrograms showing phylogenetic relationships and relative similarity among examined strains were obtained. It was also tested susceptibility of these strains to different antibiotics. All strains were resistant to nalidixic acid and more or less susceptible to tetracycline, ampicillin, streptomycin and chloramphenicol. The results clearly indicated existence of considerable diversity of autochthonous L. plantarum strains which is visible through their molecular characteristics as genomic and protein polymorphism.

• Keywords: Lactobacillus plantarum; molekularna diferencijacija; WCP/SDS-PAGE; PFGE; klasterska analiza
• Keywords (english): Lactobacillus plantarum; molecular differentiation; WCP/SDS-PAGE; PFGE; cluster analysis
• Language: croatian
• URN:NBN: urn:nbn:hr:159:051516
• 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: 2017-05-17 12:46:31