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利用本地Blast,将灰盖鬼伞的高速泳动族蛋白(high-mobility-group box,HMGB)编码基因exp与草菇PYd21基因组进行比对,找到一个exp的同源基因vv-exp。克隆测序PYd21与PYd15两单孢菌株中vv-exp基因并进行比对,结果表明2个菌株的vv-exp基因序列一致。结合本实验室草菇转录组数据对vv-exp基因结构进行分析,结果显示此基因全长为1 937bp,共含3个内含子;开放阅读框长度为1 773bp,编码590个氨基酸。预测的氨基酸序列生物信息学分析结果表明,此基因编码的蛋白含有两个HMG-box结构域。通过实时荧光定量PCR(quantitative Real-time PCR,q RT-PCR)方法分析了vv-exp基因在草菇不同生长阶段以及不同部位的表达量,结果显示该基因在原基表达量较高,表明此基因可能与草菇子实体的形成有关;另外,在成熟期菌盖中vv-exp表达量比原基上调达到3倍以上,且显著高于伸长期菌盖,由此推断该基因参与调控草菇子实体开伞。
Using local Blast, the high-mobility-group box (HMGB) coding gene exp was compared with that of the grass mushroom PYd21 genome to find a homologous gene exp of vv-exp. The vv-exp gene was cloned and sequenced in two PsD21 and PYd15 strains. The results showed that the two strains had the same vv-exp gene sequence. The vv-exp gene structure was analyzed based on the data from the Ophiopogon japonicus transcriptome. The results showed that this gene was 1 937 bp in length and contained 3 introns. The open reading frame was 1 773 bp in length and encoded 590 amino acids. Bioinformatics analysis of the predicted amino acid sequence indicated that the protein encoded by this gene contained two HMG-box domains. The expression of vv-exp gene in different growth stages and different parts of Volvariella volvacea was analyzed by quantitative real-time PCR (q RT-PCR). The results showed that the expression level of vv-exp gene was higher in the primordia, The gene may be related to the formation of Agrobacterium fruiting bodies. In addition, the expression level of vv-exp in the cap was three times higher than that in the primordial stage, and was significantly higher than that in the elongating stage. Thus, it was concluded that the gene involved in the regulation of grass Mushrooms body umbrella.