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

作者：丁少军; J.A.BUSWELLvolvariellavolvaceaxylanasecdna

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 xynl was 1287 bp in length, including 3′and 5′-non-coding region. The xynl 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 xynl with other xylanases showed that the homology with family-10 xylanases from Agaricus bisporus xyll, AspergiUus sojae xynl, Aspergillus kawachii xynA, Fusarium oxysporumf, sp xyl3 was 64% ,55% ,52% ,55% , respectively.

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