草菇钙调磷酸酶催化亚基(Vv-CNA)的序列与表达

【目的】分析草菇钙调磷酸酶催化亚基(Vv-CNA)的序列与表达,为进一步探究草菇生长发育及子实体开伞分子调控机理提供参考。【方法】通过生物信息学分析确定Vv-CNA基因的组成及保守结构,将ORF Finder预测的草菇CNA氨基酸序列与从NCBI下载的其他真菌CNA氨基酸序列进行比对,并通过荧光定量PCR检测Vv-CNA基因在草菇不同生长时期的相对表达量。【结果】Vv-CNA基因编码区为2503 bp,包含10个内含子(其中I2、I8、I9和I10存在内含子保留现象),编码610个氨基酸,Gen Bank登录号KX463514。将Vv-CNA氨基酸序列与动物(人、小鼠)、植物(拟南芥、水稻)和真菌(曲霉、酵母等)的CNA氨基酸序列进行同源比对分析,结果发现Vv-CNA蛋白有3个最保守的结构域,与密褐褶菌(Gloeophyllum trabeum)的亲缘关系最近。Vv-CNA基因在草菇子实体的蛋形期和伸长期高表达,以蛋形期的相对表达量最高。【结论】Vv-CNA基因编码钙调磷酸酶催化亚基,其高表达有利于草菇菌柄的伸长。

Sequence and expression of calcineurin catalytic subunit in Volvariella volvacea(Vv-CNA)

Objective]This research explored sequence and expression of calcineurin catalytic subunit in Volvariella volvacea(Vv-CNA), in order to provide reference for growth and development of V. volvacea and reveal the regulatory me-chanism of fruit bodies. Method]The sequence and conserved structure of Vv-CNA were analysed by bioinformatics. An-ticipated V. Volvacea CNA amino acid sequence by ORF-finder was compared with CNA amino acid sequences of other fu-gus downloaded from NCBI. Fluorescent quantitative PCR was used to detect relative expression of Vv-CNA at different growth stages of V. volvacea. Result]The coding region of Vv-CNA was 2503 bp, including 10 introns(intron retention was detected in I2, I8, I9 and I10). It encoded 610 amino acids and GenBank accession number was KX463514. Homolo-gous analysis was conducted between Vv-CNA amino acid sequence and amino acid sequences of animal(human, mouse), plant(arabidopsis, rice) and fungus(aspergillus, yeast). The results indicated that Vv-CNA contained three conserved do-mains, which were most closely related to Gloeophyllum Trabeum. Vv-CNA upregulated at egg stage and elongation stage of fruit body, and reached the peak at egg stage. Conclusion]Vv-CNA gene encodes calcineurin catalytic subunit and its up-regulated expression may be related to elongation of stipe in V. volvacea.