Genetic manipulations of Serratia marcescens with a goal of industrial application

master's thesis

• Title (croatian): Genetičke modifikacije Serratia marcescens s ciljem industrijske primjene
• Author: Marta Kozarić
• Mentor: Božidar Šantek (mentor)
• Committee member: Anamarija Štafa (član povjerenstva)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology); Zagreb
• Defense date and country: 2021-09-01, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

2,3-butanediol (2,3-BD) is one of the promising bulk chemicals that has a great potential for a variety of industries. Serratia marcescens is a Gram negative bacterium capable of producing specific meso-2,3-BD isoform. The main aim of the thesis was genetic engineering of genes connected to glucose uptake in S. marcescens. As target genes, ptsG gene and man operon were chosen and the method of choice for gene knock-out was CRISPR-Cas9. An efficient DNA transformation method for S. marcescens was established in the laboratory where electroporation showed bigger efficiency comparing to chemical competent process. Potential man operon knocked-out clones, grown on xylose as a carbon source, have been revealed but the further examination is required. PtsG gene knocked- out clones were not obtained.

Abstract (croatian)

2,3-butandiol (2,3-BD) kemikalija je s potencijalnom primjenom u raznim granama industrije. Serratia marcescens je Gram negativna bakterija koja može proizvesti specifičnu mezo-2,3-butandiol izoformu. Glavni cilj rada bio je genetički modificirati gene povezane s transportom glukoze u S. marcescens. Kao ciljane regije odabrani su gen ptsG i man operon , a za inaktivaciju gena izabrana je CRISPR- Cas9 metoda. U laboratoriju je uspostavljena učinkovita metoda genetičke transformacije bakterije S. marcescens, te je elektroporacija pokazala veću efikasnost u usporedbi s kemijski kompetentnim postupkom. U konačnici, dobiveni su klonovi u kojima je došlo do moguće inaktivacije man operona, izrasli na ksilozi kao izvoru ugljika, međutim potrebna su daljnja istraživanja. Klonovi u kojima je došlo do inaktivacije gena ptsG nisu dobiveni.

• Keywords: genetic transformation; Serratia marcescens; CRISPR-Cas9; gene knock-out
• Keywords (english): genetička transformacija; Serratia marcescens; CRISPR-Cas 9; inaktivacija gena
• Language: english
• URN:NBN: urn:nbn:hr:159:274300
• Study programme: Title: Molecular Biotechnology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2021-10-06 12:33:41