Sequence-based factors influencing the expression of heterologous genes in the yeast Pichia pastoris: A comparative view on 79 human genes.

Item Type:	Article
Title:	Sequence-based factors influencing the expression of heterologous genes in the yeast Pichia pastoris: A comparative view on 79 human genes.
Creators Name:	Boettner, M. and Steffens, C. and von Mering, C. and Bork, P. and Stahl, U. and Lang, C.
Abstract:	High yield expression of heterologous proteins is usually a matter of "trial and error". In the search of parameters with a major impact on expression, we have applied a comparative analysis to 79 different human cDNAs expressed in Pichia pastoris. The cDNAs were cloned in an expression vector for intracellular expression and recombinant protein expression was monitored in a standardized procedure and classified with respect to the expression level. Of all sequence-based parameters with a possible influence on the expression level, more than 10 were analysed. Three of those factors proved to have a statistically significant association with the expression level. Low abundance of AT-rich regions in the cDNA associates with a high expression level. A comparatively high isoelectric point of the recombinant protein associates with failure of expression and, finally, the occurrence of a protein homologue in yeast is associated with detectable protein expression. Interestingly, some often discussed factors like codon usage or GC content did not show a significant impact on protein yield. These results could provide a basis for a knowledge-oriented optimisation of gene sequences both to increase protein yields and to help target selection and the design of high-throughput expression approaches.
Keywords:	Pichia pastoris, Heterologous protein expression, Sequence-based factors
Source:	Journal of Biotechnology
ISSN:	0168-1656
Publisher:	Elsevier
Volume:	130
Number:	1
Page Range:	1-10
Date:	31 May 2007
Official Publication:	https://doi.org/10.1016/j.jbiotec.2007.02.019

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