Cellulose binding domains (CBDs) are present in the majority of fungal cellulases and hemicellulases. Based on the conserved region of CBDs, degenerate primers were designed and used to amplify the 5′ end cDNA fragme...Cellulose binding domains (CBDs) are present in the majority of fungal cellulases and hemicellulases. Based on the conserved region of CBDs, degenerate primers were designed and used to amplify the 5′ end cDNA fragment of xylanase from Volvariella volvacea by 5′ RACE. Gene specific primer was then designed based on extreme region of 5′ end cDNA fragment and used to amplify the full length cDNA of xylanase. The cDNA of xyn 1 was 1 287 bp in length, including 3′ and 5′ non coding region. The xyn 1 cDNA contained an ORF of 1101 bp encoding 367 amino acids, in which there was a putative signal peptide with 19 amino acids. Alignment of the deduced amino acid sequence of xyn 1 with other xylanases showed that the homology with family 10 xylanases from Agaricus bisporus xyl1,Aspergillus sojae xyn1, Aspergillus kawachii xynA, Fusarium oxysporum f.sp xyl3 was 64%,55%,52%,55%, respectively.展开更多
文摘Cellulose binding domains (CBDs) are present in the majority of fungal cellulases and hemicellulases. Based on the conserved region of CBDs, degenerate primers were designed and used to amplify the 5′ end cDNA fragment of xylanase from Volvariella volvacea by 5′ RACE. Gene specific primer was then designed based on extreme region of 5′ end cDNA fragment and used to amplify the full length cDNA of xylanase. The cDNA of xyn 1 was 1 287 bp in length, including 3′ and 5′ non coding region. The xyn 1 cDNA contained an ORF of 1101 bp encoding 367 amino acids, in which there was a putative signal peptide with 19 amino acids. Alignment of the deduced amino acid sequence of xyn 1 with other xylanases showed that the homology with family 10 xylanases from Agaricus bisporus xyl1,Aspergillus sojae xyn1, Aspergillus kawachii xynA, Fusarium oxysporum f.sp xyl3 was 64%,55%,52%,55%, respectively.