Your browser does not support JavaScript!

Graduate theses

Current Record: 20 of 233

Back to Results Previous page
Next page
Add to Basket
[Add to Basket]
Title Δημιουργία στελεχών Pichia pastoris για την ετερόλογη έκφραση πρωτεϊνών γλυκιάς γεύσης
Author Βουγιατζή, Αικατερίνη Σταυρούλα
Thesis advisor Παυλίδης, Ιωάννης
Abstract One of the most popular heterologous protein expression systems is the methylotrophic yeast Pichia pastoris. Using the yeast metabolic pathway for the utilization of methanol (Methanol Utilization pathway, MUt), and specifically the characteristic of the alcohol oxidase (AOX) promoter being activated only when methanol is present in the environment of the cells, the tightly controlled expression of heterologous proteins can take place. Being a eukaryotic microorganism, P. pastoris offers a more complex protein expression system than bacterial hosts, such as E. coli, while being a yeast, it is a cheaper and less time-consuming system than mammalian cells. The objective of this project was the preparation of Pichia pastoris strains for the heterologous expression of five hypothetical and two known sweet proteins. The first step of the process was to transform the yeast strain GS115 with linearized plasmid vector pPIC9K. In the following steps, growth experiments were performed on the transformed cells under specific conditions in solid-state cultures. The MUts or MUt+ phenotype of the colonies was determined, as well as their genotype by detecting the desired genes in the yeast genome using polymerase chain reaction (PCR). Finally, cultures were prepared for the expression of the proteins of interest and affinity chromatography was used for their purification. The expression of the two proteins Mab II and MNEI was observed through the electrophoresis under denaturing conditions (SDS-PAGE), but in low yields.
Language Greek
Issue date 2022-11-24
Collection   School/Department--School of Sciences and Engineering--Department of Chemistry--Graduate theses
  Type of Work--Graduate theses
Permanent Link https://elocus.lib.uoc.gr//dlib/7/d/6/metadata-dlib-1667896608-517701-9912.tkl Bookmark and Share
Views 218

Digital Documents
No preview available

Download document
View document
Views : 7