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利用真菌纤维素结合域(CBD)保守性序列进行草菇木聚糖酶cDNA的克隆 被引量:6

Cloning and Sequencing of Xylanase cDNA from Volvariella volvacea Using Conserved Sequences in Cellulose-Binding Domain
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摘要 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. 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.
出处 《中国生物化学与分子生物学报》 CAS CSCD 北大核心 2004年第3期408-412,共5页 Chinese Journal of Biochemistry and Molecular Biology
关键词 Volvariella volvacea cellulose-binding domain(CBD) xylanase CDNA Volvariella volvacea, cellulose-binding domain(CBD), xylanase, cDNA
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