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Nucleic Acids Research, 1993, Vol. 21, No. 21 4923-4928
© 1993

MOLECULAR BIOLOGY

Rational design and PCR-based synthesis of an artificial Schizophyllum commune xylanase gene

Roger W. Graham, Tom Atkinson1, D.G. Kilburn, R.C. Miller, Jr and R.A.J Warren

Department of Microbiology, University of British Columbia Vancouver, BC V6T 1W5 1ID Biomedical Corporation Vancouver V6E 2K3, Canada

Received July 19, 1993. Revised September 12, 1993. Accepted September 12, 1993.

A synthetic gene encoding the Schizophyllum commune xylanase XynA was constructed by a novel PCR-based procedure. Threelong oligonucleotides were synthesized and used in combination with flanking PCR primers to generate a 607 base pair gene which contained 31 unique locations for restriction enzyme cleavage. The amino acid sequence was tailored for expression in Eschericia coli by using only those codons found in highly expressed E.coli genes. Theavailability of the gene will facilitate analysis of the structure and function of this and other ß-(1,4) xylanases.


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