prikaz prve stranice dokumenta Izoelektrično fokusiranje proteina stanične stjenke kvasca Saccharomyces cerevisiae
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master's thesis
Izoelektrično fokusiranje proteina stanične stjenke kvasca Saccharomyces cerevisiae
2009. urn:nbn:hr:109:677405
Gradvol, Vedran
Josip Juraj Strossmayer University of Osijek
FACULTY OF FOOD TECHNOLOGY
Department of Applied Chemistry and Ecology
Sub-department of Biochemistry and Toxicology

Cite this document

Gradvol, V. (2009). Izoelektrično fokusiranje proteina stanične stjenke kvasca Saccharomyces cerevisiae (Master's thesis). Osijek: Josip Juraj Strossmayer University of Osijek, FACULTY OF FOOD TECHNOLOGY. Retrieved from https://urn.nsk.hr/urn:nbn:hr:109:677405

Gradvol, Vedran. "Izoelektrično fokusiranje proteina stanične stjenke kvasca Saccharomyces cerevisiae." Master's thesis, Josip Juraj Strossmayer University of Osijek, FACULTY OF FOOD TECHNOLOGY, 2009. https://urn.nsk.hr/urn:nbn:hr:109:677405

Gradvol, Vedran. "Izoelektrično fokusiranje proteina stanične stjenke kvasca Saccharomyces cerevisiae." Master's thesis, Josip Juraj Strossmayer University of Osijek, FACULTY OF FOOD TECHNOLOGY, 2009. https://urn.nsk.hr/urn:nbn:hr:109:677405

Gradvol, V. (2009). 'Izoelektrično fokusiranje proteina stanične stjenke kvasca Saccharomyces cerevisiae', Master's thesis, Josip Juraj Strossmayer University of Osijek, FACULTY OF FOOD TECHNOLOGY, accessed 13 April 2024, https://urn.nsk.hr/urn:nbn:hr:109:677405

Gradvol V. Izoelektrično fokusiranje proteina stanične stjenke kvasca Saccharomyces cerevisiae [Master's thesis]. Osijek: Josip Juraj Strossmayer University of Osijek, FACULTY OF FOOD TECHNOLOGY; 2009 [cited 2024 April 13] Available at: https://urn.nsk.hr/urn:nbn:hr:109:677405

V. Gradvol, "Izoelektrično fokusiranje proteina stanične stjenke kvasca Saccharomyces cerevisiae", Master's thesis, Josip Juraj Strossmayer University of Osijek, FACULTY OF FOOD TECHNOLOGY, Osijek, 2009. Available at: https://urn.nsk.hr/urn:nbn:hr:109:677405

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Metadata
TitleIzoelektrično fokusiranje proteina stanične stjenke kvasca Saccharomyces cerevisiae
Title (english)Isoelectric focusing of Saccharomyces cerevisiae cell wall proteins
AuthorVedran Gradvol
MentorIvica Strelec (mentor)
Committee memberMarija Halt (član povjerenstva)
MBZ: 76414
Committee memberIvica strelec (član povjerenstva)
Committee memberVladimir Mrša (član povjerenstva)
MBZ: 39525
Committee memberHrvoje Pavlović (član povjerenstva)
GranterJosip Juraj Strossmayer University of Osijek
FACULTY OF FOOD TECHNOLOGY
(Department of Applied Chemistry and Ecology)
(Sub-department of Biochemistry and Toxicology)
Osijek
Defense date and country2009-09-30, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Food Technology
Engineering
Abstract
U ovom radu su nekovalentno i kovalentno vezani proteini stanične stijenke različitih mutanata kvasca koji su se razlikovali u proteinima stanične stijenke ekstrahirani pomoću 20 mM NaOH i 20 IU/μL -1,3-glukanaze, nakon čega je uslijedilo izoelektrično fokusiranje u gradijentu pH 3-10, te nespecifična i specifična detekcija proteina. Nespecifična detekcija proteina je provedena bojanjem srebrom, a specifična detekcija imunokemijski nakon kapilarno-difuzijskog prijenosa proteina na PVDF membranu. Nespecifičnom detekcijom proteina htjelo se ustanoviti da li postoje razlike u količini ekstrahiranih proteina i rasporedu proteinskih vrpci različitih mutanata kvasca, a imunokemijskom detekcijom se htjelo utvrditi prisustvo nekovalentno vezanih proteina Scw4p, Scw10p, Scw11p i Bgl2p označenih hemaglutininskim epitopom. Iz provedenog izoelektričnog fokusiranja, odnosno nakon nespecificne detekcije bojanjem srebrom mogu se uočiti razlike u ekspresiji proteina pojedinih mutanata kvasca u vidu razlika u intenzitetu pojedinih proteinskih vrpci, dok istovremeno nema evidentnih razlika u samom rasporedu proteinskih vrpci. Prilikom razdjeljivanja proteina došlo je i do taloženja proteina na mjestu nanošenja što se otklonilo dodatkom Tritona X-100 u uzorke ekstrahiranih proteina. Imunokemijskom detekcijom pokazano je nespecifično vezanje proteina što je vjerojatno uzrokovano pucanjem stanica kvasca prilikom ekstrakcije i nespecifične reakcije unutarstaničnih proteina sa HAprotutijelima.
Abstract (english)
Covalently and noncovalently linked cell wall proteins from different yeast protein mutants were extracted with 20 mM NaOH and 20 IU/μL -1,3-glucanase. Extracted proteins were separated by isoelectric focusing in a pH gradient of 3-10, and subsequently nonspecific and specific detection was carried out. Nonspecific detection was achieved by silver staining and specific detection was achieved immunochemically after the transfer of proteins to a PVDF membrane. The goal of nonspecific detection was to determine the differences in the quantity of extracted proteins and/or position of protein bands for the different yeast mutants, and the goal of specific detection was to determine the presence of noncovalently linked proteins Scw4p, Scw10p, Scw11p and Bgl2p tagged by haemaglutinin. Silver stained gels showed differences in protein expression of different yeast mutants that can be seen as differences of protein band intensities, but there were no evident differences in protein band positions of different yeast mutants. Protein precipitation during sample entry was omitted by addition of Triton X-100 in all samples of extracted proteins that increased protein solubility. Immunochemical detection showed nonspecific reaction of HA-antibodies with yeast proteins which was probably caused by the release of intercellular proteins after cell breakage during extraction.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:109:677405
ProjectNumber: 113-1130473-0334
Title: Sinergističke smjese u antifungalnoj i antimikotoksigenoj zaštiti hrane
Title: Synergistic mixtures in antifungal and antimycotoxigenic food protection
Leader: Tomislav Klapec
Jurisdiction: Croatia
Funder: MZOS
Funding stream: ZP
ProjectNumber: 113-1130475-2722
Title: Unapređenje kakvoće, trajnosti i funkcionalnosti proizvoda na bazi pšenice
Title: Improvement of quality, shelf life and functionality of wheat-based products
Leader: Daliborka Koceva Komlenić
Jurisdiction: Croatia
Funder: MZOS
Funding stream: ZP
Study programmeTitle: Food Technology and Process Engineering
Study programme type: university
Study level: graduate
Academic / professional title: diplomirani inženjer prehrambene tehnologije i procesnog inženjerstva (diplomirani inženjer prehrambene tehnologije i procesnog inženjerstva)
Type of resourceText
File originBorn digital
Access conditionsAccess restricted to students and staff of home institution
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Created on2015-12-02 15:26:54