目的 克隆桑黄菌丝甾醇C14还原酶（ERG24）基因全长，并进行生物信息学及表达模式的初步分析。方法 基于桑黄菌丝转录组数据设计PlERG24基因引物，采用PCR技术获得其cDNA全长序列，并通过Ex PASy等在线分析软件对其进行生物信息学分析，同时利用实时荧光定量PCR技术分析该基因的表达模式。结果 从桑黄菌丝中克隆到1 412 bp的PlERG24基因，开放阅读框长度为1 326 bp，编码441个氨基酸，理论相对分子质量为49 358.61，等电点为5.28，为不含信号肽的疏水性蛋白，推测定位于质膜，具有6个磷酸化位点；系统进化树分析表明，该序列与暴马桑黄的ERG24基因同源性最高。表达模式分析表明，在桑黄菌丝生长的一个周期内，PlERG24基因表达在25 d时达到最高6.36。结论 获得了PlERG24基因全长序列，为进一步研究该基因的功能以及利用基因工程等手段改良麦角甾醇生物合成途径奠定基础。
Objective To clone the gene full length of ergosterol C14 reductase (ERG24) in Phellinus linteus and analyze its bioinformatics and expression pattern. Methods The primers of PlERG24 were designed according to the transcription sequence of P. linteus, the cDNA full-length sequence of PlERG24 was obtained by PCR, its bioinformatics was analyzed by Ex PASy and other online analysis software, and its expression pattern in mycelia of P. linteus was analyzed by real-time fluorescence quantitative PCR. Results The full-length cDNA of PlERG24 gene was 1 412 bp, which encoding a protein of 441 amino acids with a predicted molecular weight of 49 358.61 and isoelectric point of 5.28; ERG24 protein was a hydrophobic protein without signal peptide, which was presumably located in the plasma membrane with six phosphorylation sites. Phylogenetic tree analysis indicated that amino acid sequences of ERG24 in P. linteus were genetically closely related to ERG24 in Sanghuangporus baumii. The qRT-PCR results showed that gene expression of PlERG24 reached the highest level of 6.36 at 25d during the growth cycle of mycelia in P. linteus. Conclusion The full length of PlERG24 gene was obtained, which lays a foundation for further studies on gene function and genetic regulatory mechanism of ergosterol biosynthesis.