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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。
Relying on the mushroom genome and transcriptome data, one glutathione peroxidase (GPx) gene was cloned and named as vvgpx01. The results of ZOOM software showed that the gene was 1 313 bp in length and contained 4 exons and 3 introns, encoding 228 amino acids. The 5 ’untranslated region was 12 bp in length and the 3’ untranslated region Which was 452 bp in length. Among them, 3 introns showed retention during RNA processing. The results of bioinformatics analysis showed that the GPx01 sequence of grass mushroom had a complete glutathione peroxidase conserved domain. The phylogenetic tree showed that GPx of Basidiomycetes and Ascomycetes clustered in two different branches respectively, Agaricus GPx01 is closely related to Agaricus bisporus and collects on one branch with other fungi of the order Agaric. In addition, amino acid sequences produced by intron retention all showed varying degrees of loss of domain, which led to the inactivation of glutathione peroxidase. Expression profiling results showed that: vv-gpx01 expression in the entire growth stage of mushroom, but there are differences in expression levels. Speculated that the grass mushroom GPx01 may produce different transcripts by alternative splicing, by regulating the expression of different transcripts to regulate gene function domain translation and spatial configuration, and ultimately the regulation of enzyme activity.