Abstract
Four vectors were constructed for high-level expression of heterologous proteins with high copy number and mitotic stability in Hansenula polymorpha. All of them contained the conserved H. polymorpha-derived ribosomal DNA (rDNA) sequence for targeting and the geneticin (G418) resistance gene as a selection marker. A strong inducible promoter, formate dehydrogenase (FMD) promoter from H. polymorpha, was used to drive the expression of heterologous genes; the formate dehydrogenase terminator of H. polymorpha was used as the transcription termination region. A modified green fluorescent protein (mGFP) and firefly luciferase protein (Luc) were used as the marker to evaluate the efficacy of these vectors. Using Southern blotting analysis, 2–30 copies of these vectors were integrated into rDNA loci. These results demonstrated that all the four vectors could be used as candidates for expression of desired proteins in H. polymorpha.
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References
H Cox D Mead P Sudbery RM Eland I Mannazzu L Evans (2000) ArticleTitleConstitutive expression of recombinant proteins in the methylotrophic yeast Hansenula polymorpha using the PMA1 promoter Yeast 16 1191–1203
A Degelmann F Muller H Sieber V Jenzelewski M Suckow AW Strasser G Gellissen (2002) ArticleTitleStrain and process development for the production of human cytokines in Hansenula polymorpha FEMS Yeast Res. 2 349–361
KN Faber P Haima W Harder M Veenhuis G Ab (1994) ArticleTitleHighly-efficient electrotransformation of the yeast Hansenula polymorpha Curr. Genet. 25 305–310
AJ Fellinger JM Verbakel RA Veale PE Sudbery IJ Bom N Overbeeke CT Verrips (1991) ArticleTitleExpression of the alpha-galactosidase from Cyamopsis tetragonoloba (guar) by Hansenula polymorpha Yeast 7 463–473
JW Harger JD Dinman (2003) ArticleTitleAn in vivo dual-luciferase assay system for studying translational recoding in the yeast Saccharomyces cerevisiae RNA 9 1019–1024
G Gellissen ZA Janowicz A Merckelbach M Piontek P Keup U Weydemann CP Hollenberg AW Strasser (1991) ArticleTitleHeterologous gene expression in Hansenula polymorpha: efficient secretion of glucoamylase Biotechnology 9 291–295
ZA Janowicz K Melber A Merckelbach E Jacobs N Harford M Comberbach CP Hollenberg (1991) ArticleTitleSimultaneous expression of the S and L surface antigens of hepatitis B, and formation of mixed particles in the methylotrophic yeast, Hansenula polymorpha Yeast 7 431–443
AF Mayer K Hellmuth H Schlieker R Lopez-Ulibarri S Oertel U Dahlems AW Strasser AP Loon Particlevan (1999) ArticleTitleAn expression system matures: a highly efficient and cost-effective process for phytase production by recombinant strains of Hansenula polymorpha Biotechnol. Bioeng. 63 373–381
JH Sohn ES Choi HA Kang JS Rhee SK Rhee (1999) ArticleTitleA family of telomere-associated autonomously replicating sequences and their functions in targeted recombination in Hansenula polymorpha DL-1 J. Bacteriol. 181 1005–1013
H Song Y Li W Fang Y Geng X Wang M Wang B Qiu (2003) ArticleTitleDevelopment of a set of expression vectors in Hansenula polymorpha Biotechnol. Lett. 25 1999–2006
T Wartmann G Kunze (2000) ArticleTitleGenetic transformation and biotechnological application of the yeast Arxula adeninivorans Appl. Microbiol. Biotechnol. 54 619–624
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Liu, Y., Li, Y., Liu, L. et al. Design of Vectors for Efficient Integration and Transformation in Hansenula polymorpha. Biotechnol Lett 27, 1529–1534 (2005). https://doi.org/10.1007/s10529-005-1469-7
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DOI: https://doi.org/10.1007/s10529-005-1469-7