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

Paper ID Volume ID Publish Year Pages File Format Full-Text
25289 43565 2007 10 PDF Available
Title
Sequence-based factors influencing the expression of heterologous genes in the yeast Pichia pastoris—A comparative view on 79 human genes
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
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Sequence-based factors influencing the expression of heterologous genes in the yeast Pichia pastoris—A comparative view on 79 human genes
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 130, Issue 1, 31 May 2007, Pages 1–10
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us