Multiple chromosomal gene integration for production of pharmaceutical proteins in S. cerevisiae

Malene Jensen, Uffe Hasbro Mortensen, Nina Gunnarsson, Tomas Strucko, Line Due Baron

Research output: Contribution to journalConference abstract in journalResearchpeer-review

Abstract

When studying protein folding and secretion the general conception is that all cells in a population express an equal amount of protein. Recent work has shown that expression levels vary greatly in cell populations which express proteins on plasmids. Hence a yeast expression platform has been developed at the Department of Systems Biology, DTU. The platform offers the opportunity to express genes on the chromosome in 1 to 10 copies. A comparison between the expression of CFP and RFP by the platform and by plasmids reveals the problems of plasmid expression. FACS analyses of two cell populations, expressing CFP and RFP on the separate plasmids or expressing CFP and RFP using the yeast expression platform shows expression varies greatly in a cell population based on plasmid expression compared to the yeast expression platform. When expressed on plasmids a few cells are high performers on both proteins but the largest fraction of cells is actually not expressing either of the proteins. The yeast expression platform is developed to facilitate stable expression of integrated genes. The integration sites are separated by essential genes which ensure that the integrated genes are not lost by recombination. An amplification method has been developed for the platform which enables fast integration of genes. Future perspectives involve exploring the capabilities of the platform for recombinant protein production including performance and stability studies.
Original languageEnglish
JournalNew Biotechnology
Volume31
Issue numberSupplement
Pages (from-to)S192
ISSN1871-6784
DOIs
Publication statusPublished - 2014
Event16th European Congress on Biotechnology - Edinburgh, United Kingdom
Duration: 13 Jul 201416 Jul 2014
Conference number: 16

Conference

Conference16th European Congress on Biotechnology
Number16
CountryUnited Kingdom
CityEdinburgh
Period13/07/201416/07/2014

Bibliographical note

Copyright © 2014 Published by Elsevier B.V.

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