草菇谷胱甘肽过氧化物酶编码基因vv-gpx01的序列特征及表达分析

Sequence Characteristics and Expression Analysis of a Glutathione Peroxidase Gene vv-gpx01 from Volvariella volvacea

依托草菇基因组及转录组数据,克隆得到1个谷胱甘肽过氧化物酶(GPx)编码基因,命名为vvgpx01。ZOOM软件的分析结果表明:该基因全长1 313 bp,含有4个外显子、3个内含子,编码228个氨基酸;5'端非翻译区长为12 bp,3'端非翻译区为452 bp;其中3个内含子在RNA加工过程中均存在保留现象。生物信息学分析结果表明:草菇的GPx01序列具备完整的谷胱甘肽过氧化物酶保守结构域;系统发育树显示,担子菌与子囊菌的GPx分别聚在了2个不同的分支上,草菇GPx01与双孢蘑菇亲缘关系较近并与其他伞菌目的真菌聚在1个分支上。另外,由内含子保留所产生的氨基酸序列均出现了不同程度的功能域缺失,这将导致谷胱甘肽过氧化物酶的失活。表达谱分析结果表明:vv-gpx01在整个草菇生长阶段都有表达,但表达量存在差异。推测,草菇GPx01可能通过可变剪接产生不同的转录本,通过调节不同转录本的表达水平来调控基因功能域的翻译水平及空间构型,最终实现酶活性的调节。

Based on genome and transcriptome data of Volvariella volvacea, a gene encoding gluta？thione peroxidase （ GPx） was cloned, and named as vv？gpx01. The results analyzed by ZOOM soft？ware showed that vv？gpx01 contained 1 313 bp, encoding a polypeptide of 228 amino acids. vv？gpx01 gene contained 4 exons and 3 introns, 5′UTR contained 12 bp, 3′UTR contained 452 bp. And three intron retention alternative splicing sites were identified during RNA processing. The bioinformatics analysis showed GPx01 of Volvariella volvacea had a complete conserved domain of glutathione peroxidase. The analysis of phylogenetic tree showed that glutathione peroxidase of basid？iomycetes and ascomycetes were gathered in two different branches, GPx01 of Volvariella volvacea showed the highest homology with Agaricus bisporus and was gathered in a branch with other Agari？cales sequences. Furthermore, the amino acid sequences generated from the intron retention showed the missing of GSH peroxidase domain in different levels, which probably caused the deactivation of glutathione peroxidase. The analysis of digital gene expression profiles showed that vv？gpx01 in Volvariella volvacea was expressed during all the growth stages, but the expression value was differ？ent. It suggested that the alternative splicing probably produced different transcripts of GPx01, con？trolling the translation of functional domain and spatial configuration by regulating the expression lev？els of different transcripts, and ultimately regulating the enzyme activity.