摘要
以阿魏侧耳(Pleurotus ferulae lanzi)CGMCC 5.774为实验材料,采用同源基因克隆方法克隆漆酶(Laccase,Lacc)基因。根据刺芹侧耳漆酶基因序列(AM774000.1、DQ234990.1)设计阿魏侧耳漆酶基因扩增引物,以阿魏侧耳c DNA为模板克隆漆酶基因CDS区序列,获得2条阿魏侧耳漆酶基因序列分别命名为PFLacc A(Genbank登录号KP246838)和PFLacc B(Genbank登录号KP246839)。PFLacc A(KP246838)和PFLacc B CDS(KP246839)序列长度均为1602 bp,与刺芹侧耳漆酶全CDS区相似性为97%,阿魏侧耳漆酶基因与刺芹侧耳漆酶基因具有高度的同源性。PFLacc A CDS和PFLacc B CDS序列均为全长的开放阅读框ORF,编码蛋白质长度为533 aa。PFLacc A蛋白和PFLacc B蛋白均包含长度为20 aa的信号肽和Cu-oxidase_3、Cu-oxidase、Cu-oxidase_2 3个结构域;Predict Protein对PFLacc A和PFLacc B二级结构进行分析,并通过SWISS-MODEL预测了该酶的三维结构。
Laccase gene was cloned from experiment materials of Pleurotus ferulae lanzi（CGMCC 5.0774）. Pleurotus ferulae laccase gene amplifi cation primers was designed according to the sequence of Pleurotus eryngii laccase gene（AM774000.1, DQ234990.1）. The region of laccase CDS was cloned according to the template of Pleurotus ferulae c DNA. Both of the sequence length of PFLacc A CDS（KP246838） and PFLacc B CDS（KP246839） were 1602 bp, PFLacc A CDS and PFLacc B CDS were both full-length sequences of the open reading frame ORF, encoding a protein length of 533 aa. Both proteins were contain an amino-terminal signal peptide with the size of 20 amino acid residues and the domain of Cu-oxidase3, Cu-oxidase, Cu-oxidase2 for PFLacc A and PFLacc B. Laccase Gene from Pleurotus ferulae has high homology with laccase of Pleurotus eryngii. Furthermore analysised secondary structures of PFLacc A and PFLacc B, and predicted the three-dimensional structure of the enzymes by SWISSMODEL.
出处
《食品科技》
CAS
北大核心
2015年第5期14-20,共7页
Food Science and Technology
基金
国家自然科学基金项目(31160312)
关键词
阿魏侧耳
漆酶
基因克隆
Pleurotus ferulae lanzi
laccase
gene cloning
