【摘要】 以白黄侧耳（Pleurotus cornucopiae）为材料，在全基因组范围内检索并克隆酪氨酸酶（tyrosinase,TYR）基因，利用生物信息学方法分析基因结构、染色体定位、顺式作用元件、保守基序、三维结构、信号肽、亚细胞定位和理化性质，构建系统进化树，分析保守结构域，采用荧光定量PCR分析TYR基因在子实体不同发育阶段和不同部位的相对表达量。结果表明：全基因组范围内共克隆得到4个基因，分别命名为PcTYR1、PcTYR2、PcTYR3和PcTYR4，分别分布于2条不同染色体上，均含有与激素反应及非生物胁迫应答相关的顺式作用元件，且三维结构相似，均具有两个高度保守的铜离子结合位点，在保守基序、信号肽、亚细胞定位和理化性质方面存在差异；PcTYR1的相对表达量在整个发育过程中先增加后降低，由菌丝体的0.96±0.04增加至幼菇的156.78±5.09，随后下降至孢子的2.49±0.46,PcTYR1和PcTYR4在原基和幼菇中的相对表达量大于PcTYR2和PcTYR3,PcTYR基因家族在子实体的不同部位中均有表达，PcTYR1、PcTYR2和PcTYR4在菌盖中的相对表达量均显著高于菌褶和菌柄。更多还原

【Abstract】 Tyrosinase(TYR) genes were identified and cloned from the whole genome of Pleurotus cornucopiae.The identified PcTYRs and their protein products were analyzed by bioinformatics tools for gene structure,chromosomal mapping, cis-acting elements, conserved motifs, 3D protein structure, signal peptide, subcellular localization and physiochemical properties. Phylogenetic tree was constructed based on amino acid sequences of TYRs, and conserved domains were analyzed. The relative expression of PcTYRs in different developmental stages and parts of fruiting bodies were determined by fluorescence quantitative PCR. The results showed that a total of four tyrosinase genes, PcTYR1, PcTYR2, PcTYR3 and PcTYR4, were cloned from the whole genome of P. cornucopiae. They are distributed on two different chromosomes. All four genes contain cis-acting elements related to hormone response and abiotic stress response, and two highly conserved copper ion binding sites.Regardless of similar 3D protein structures, there are differences in conserved motifs, signal peptide, subcellular localization and protein properties between the PcTYRs. During the entire course of fruiting body development,the relative expression level of PcTYR1 initially increased from 0.96 in mycelium to 156.78 in young fruiting bodies, and then decreased to 2.49 in spores. The relative expression levels of PcTYR1 and PcTYR4 were higher than that of PcTYR2 and PcTYR3 in primordia and young fruiting bodies. The expression of PcTYR1-PcTYR4was detected in all parts of fruiting body, and the expression of PcTYR1, PcTYR2 and PcTYR4 was higher in pileus than in gill and stipe.更多还原