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Internal ribosome entry site mediates protein synthesis in yeast Pichia pastoris
- Source :
- Biotechnology letters. 34(5)
- Publication Year :
- 2012
-
Abstract
- The imitation of translation, as mediated by internal ribosome entry sites, has not yet been reported in Pichia pastoris. An IRES element from Saccharomyces cerevisiae was demonstrated to direct the translation of a dicistronic mRNA in P. pastoris. The 5′-untranslated region of GPR1 mRNA, termed GPR, was cloned into a dual reporter construct containing an upstream Rhizomucor miehei lipase (RML) and a downstream β-galactosidase gene (lacZ) from Escherichia coli BL21. After being transformed into P. pastoris, the RML gene and lacZ were simultaneously expressed. The possibility of DNA rearrangement, spurious splicing, or cryptic promoter in the GPR sequence were eliminated, indicating that expression of a second ORF was IRES-dependent. These findings strongly suggested that the IRES-dependent translation initiation mechanism is conserved in P. pastoris and provides a useful means to express multiple genes simultaneously.
- Subjects :
- Messenger RNA
biology
fungi
Bioengineering
Translation (biology)
General Medicine
Lipase
biology.organism_classification
beta-Galactosidase
Applied Microbiology and Biotechnology
Molecular biology
Pichia
Pichia pastoris
Cell biology
Internal ribosome entry site
Eukaryotic translation
Genes, Reporter
Protein Biosynthesis
RNA splicing
Protein biosynthesis
Escherichia coli
Rhizomucor
Gene
Ribosomes
Biotechnology
Subjects
Details
- ISSN :
- 15736776
- Volume :
- 34
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- Biotechnology letters
- Accession number :
- edsair.doi.dedup.....66f6789e032a34b1313a0fe61d4b3253