Codon optimization of xylanase genexynBfrom the thermophilic bacteriumDictyoglomus thermophilumfor expression in the filamentous fungusTrichoderma reesei
Open Access
- 1 September 2000
- journal article
- Published by Oxford University Press (OUP) in FEMS Microbiology Letters
- Vol. 190 (1) , 13-19
- https://doi.org/10.1111/j.1574-6968.2000.tb09255.x
Abstract
The catalytic domain of the xynB (xylanase) gene from the thermophilic bacterium Dictyoglomus thermophilum was reconstructed by PCR to match the codon preference of Trichoderma reesei. The 0.6-kb DNA fragment encoding the enzyme was first amplified by primer extension with a mixture of eight overlapping oligonucleotides, followed by PCR with outside primers containing restriction enzyme sites for directional cloning into Escherichia coli and T. reesei vectors. The synthetic gene was expressed in both organisms, producing a clearing halo around transformant colonies in plate assay utilizing an overlay of oat spelts xylan. Effective transcription of xynB in T. reesei was obtained after changing 20 codons.Keywords
This publication has 14 references indexed in Scilit:
- Molecular diversity of thermophilic cellulolytic and hemicellulolytic bacteriaFEMS Microbiology Ecology, 1999
- Gene Synthesis by a LCR-Based Approach: High-Level Production of Leptin-L54 Using Synthetic Gene inEscherichia coliBiochemical and Biophysical Research Communications, 1998
- Efficient production of secreted proteins by Aspergillus : progress, limitations and prospectsApplied Microbiology and Biotechnology, 1997
- Families and Functions of Novel Thermophilic Xylanases in the Facilitated Bleaching of PulpPublished by American Chemical Society (ACS) ,1996
- Rational design and PCR-based synthesis of an artificialSchizophyllum communexylanase geneNucleic Acids Research, 1993
- Foreign gene expression in yeast: a reviewYeast, 1992
- Expression of tetanus toxin fragment C inE.coli: high level expression by removing rare codonsNucleic Acids Research, 1989
- One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution.Proceedings of the National Academy of Sciences, 1989
- Design, synthesis and expression of a human interteukin-2 gene incorporating the codon usage bias found in highly expressed Escherichia coli genesNucleic Acids Research, 1988
- A versatile transformation system for the cellulolytic filamentous fungus Trichoderma reeseiGene, 1987