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3种食用菌漆酶活性比较及其基因的克隆与序列分析
引用本文:刘腾,郭九峰,那日,薛丹.3种食用菌漆酶活性比较及其基因的克隆与序列分析[J].安徽农业科学,2013(28):11278-11285.
作者姓名:刘腾  郭九峰  那日  薛丹
作者单位:内蒙古大学自治区离子束生物工程重点实验室;
基金项目:国家自然科学基金(No.51267014)
摘    要:目的]研究食用菌漆酶基因的差异以及漆酶的活性。方法]利用编码杏鲍菇漆酶基因序列(GenbankNo.AY686700)设计特异性引物,采用PCR方法获得3种典型木腐食用菌(杏鲍菇、平菇、白灵菇)漆酶的基因序列。结果]扩增得到的片段长度均为2606bp,通过对基因序列的内含子/外显子分析,获得漆酶编码区的cDNA,该基因的开放阅读框由1596个核苷酸组成,编码一个由531个氨基酸组成的多肽。3种食用茵漆酶基因的核苷酸序列、氨基酸序列同源性均为99.81%,说明漆酶的一级结构序列具有很高的保守性。结论]序列的差异性、氨基酸跨膜区的不同及蛋白质的构象不同可能导致漆酶活性的高低。

关 键 词:食用菌  漆酶基因  克隆  酶活性  序列分析

Cloning and Sequence Analysis of the Laccase Gene and Comparison of Laccase Activity from Three Kinds of Edible Fungi
Institution:LIU Teng et al (Key Laboratory of Ion Beam Bioengineering in Inner Mongolia Autonomous Region, Hohhot, Inner Mongolia 010021 )
Abstract: Objective ] To study the differences of edible fungi laccase gene and activity of laccase. Method ] According to the laccase gene of¥ Pleurotus eryngii ¥ ( Genebank No. AY686700), specific primers were designed, three kinds of edible fungi laccase gene were obtained by using PCR. Result] The length of amplification fragment was 2 606 bp, through the intron and exon analysis of gene sequence, coding cDNA of laccase was obtained. It contains one open reading frame( ORF), which encodes for a polypeptide containing 531 amino acids. The nucleic acid homology of three edible fungi laccase genes and amino acids homology were both 99.81%. Conclusion] The sequence differ- ences, amino acid transmembrane area differences and the difference of protein conformation may cause the laccase activity of high and low.
Keywords:Edible fungi  Laccase gene  Cloning  Enzymatic activity  Sequence analysis
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