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 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 Escherichia coli by using only those codons found in highly expressed E. coli genes. The availability of the gene will facilitate analysis of the structure and function of this and other beta-(1,4) xylanases.
Graham
R W
RW
Department of Microbiology, University of British Columbia, Vancouver, Canada.
Atkinson
T
T
Kilburn
D G
DG
Miller
R C
RC
Warren
R A
RA
D001483:Base Sequence; D003001:Cloning, Molecular; D017076:Computer-Aided Design; D004274:DNA, Recombinant; D004926:Escherichia coli; D005800:Genes, Fungal; D005813:Genes, Synthetic; D006026:Glycoside Hydrolases / Q000235:genetics*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction; D012567:Schizophyllum / Q000201:enzymology* / Q000235:genetics; D043325:Xylan Endo-1,3-beta-Xylosidase
D004274:DNA, Recombinant; D006026:Glycoside Hydrolases; D043325:Xylan Endo-1,3-beta-Xylosidase