magistrsko delo
Mladen Soldat (Avtor), Kristina Sepčić (Recenzent), Uroš Petrovič (Mentor), Uroš Petrovič (Član komisije za zagovor), Matej Skočaj (Član komisije za zagovor), Kristina Sepčić (Član komisije za zagovor), Matej Butala (Član komisije za zagovor), Matej Skočaj (Komentor)

Povzetek

Egerolizini so proteini, ki so značilni predvsem za bakterije in glive, najdemo pa jih tudi pri nekaterih rastlinah in virusih. Njihova biološka vloga ni natančno znana, predvideva pa se, da naj bi imeli vlogo pri procesih kot so modulacija bakterijske sporulacije ter virulenca in razvoj plodnih telesc pri glivah. Osnovna značilnost egerolizinov je, da se vežejo na in prepoznajo specifične lipide, lipidne domene ali lipidne mešanice v membranah živih celic in v umetnih lipidnih sistemih. V skupino egerolizinov sta uvrščena tudi proteina ostreolizin A (OlyA) in nigerolizin A2 (NigA2). V okviru magistrske naloge smo želeli egerolizina OlyA in NigA2 izraziti v kvasovki Saccharomyces cerevisiae ter preveriti, ali vplivata na kopičenje nevtralnih lipidov. Egerolizina OlyA in NigA2 smo izražali s pomočjo ekspresijskega plazmida pYES2, ki vsebuje inducibilni promotor GAL1. Izražanje rekombinantnih proteinov smo inducirali z galaktozo in s točkovnim nanosom celokupnih proteinov na poliviliden-difluoridno membrano preverili prisotnost oziroma izražanje rekombinantnih egerolizinov. Količino nevtralnih lipidov v obliki lipidnih kapljic smo v rekombinantnih celicah preverili z barvilom nilsko rdeče in uporabo fluorescenčnega mikroskopa. Primerjava reprezentativnih fluorescenčnih slik kvasovke S. cerevisiae s praznim plazmidom (negativna kontrola) in rekombinantnimi plazmidi nakazuje, da egerolizina OlyA in NigA2 vplivata na kopičenje nevtralnih lipidov v obliki lipidnih kapljic v celicah kvasovke S. cerevisiae.

Ključne besede

egerolizini;ostreolizin;nigerolizin;holesterol;sfingomielin;ceramid - fosfoetanolamin;

Podatki

Jezik: Slovenski jezik
Leto izida:
Tipologija: 2.09 - Magistrsko delo
Organizacija: UL BF - Biotehniška fakulteta
Založnik: [M. Soldat]
UDK: 577.2(043.2)
COBISS: 5204047 Povezava se bo odprla v novem oknu
Št. ogledov: 760
Št. prenosov: 222
Ocena: 0 (0 glasov)
Metapodatki: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Ostali podatki

Sekundarni jezik: Angleški jezik
Sekundarni naslov: ǂThe ǂimpact of aegerolysins on the physiology of yeast Saccharomyces cerevisiae cells
Sekundarni povzetek: Aegerolysins are proteins that are mainly present bacteria and fungi, and are also found in some plants and viruses. Their biological role is not known precisely, but it is predicted that they could play a role in processes such as modulation of bacterial sporulation, virulence and fructification in fungi. The basic feature of aegerolysins is that they bind to and recognize specific lipids, lipid domains or lipid mixtures in living cell membranes and in artificial lipid systems. The proteins ostreolysin A (OlyA) and nigerolysin A2 (NigA2) are members of the aegerolysin protein group. As part of the master's thesis, we wanted to express these two aegerolysins in yeast Saccharomyces cerevisiae cells and to investigate whether their expression affects the accumulation of neutral lipids. The genes were expressed by using the plasmid pYES2 that contains an inducible GAL1 promoter. The expression of recombinant proteins was induced by galactose, and the presence of recombinant proteins was investigated by dot application of total yeast proteins to a polyvinylidene fluoride membrane. The recombinant cells were stained with Nile red dye and imaged using a fluorescence microscope. By comparing representative fluorescence images of S. cerevisiae with empty plasmid (negative control) and recombinant plasmids, we suggest that aegerolysins OlyA and NigA2 induce the accumulation of lipid droplets in baker yeast cells.
Sekundarne ključne besede: aegerolysins;ostreolysin;nigerolizin;cholesterol;sphingomyelin;ceramide - phosphoethanolamine;
Vrsta dela (COBISS): Magistrsko delo/naloga
Študijski program: 0
Konec prepovedi (OpenAIRE): 1970-01-01
Komentar na gradivo: Univ. Ljubljana, Biotehniška fak.
Strani: XII, 44 f.. [8] f. pril.
ID: 11228340