RATIONAL DESIGN AND PCR-BASED SYNTHESIS OF AN ARTIFICIAL SCHIZOPHYLLUM-COMMUNE XYLANASE GENE

A synthetic gene encoding the Schizophyllum commune xylanase XynA was constructed by a novel PCR-based procedure. Three long oligonucleotide s were synthesized and used in combination with flanking PCR primers t o generate a 607 base pair gene which contained 31 unique locations fo r restriction enzyme cleavage. The amino acid sequence was tailored fo r expression in Eschericia coli by using only those codons found in hi ghly expressed E. coli genes. The availability of the gene will facili tate analysis of the structure and function of this and other beta-(1, 4) xylanases.