旨在克隆香菇C91-3转录本Unigene 14872基因中的Pkinase结构域,并对其进行生物信息学分析。提取香菇C91-3菌丝体中总RNA,根据转录组测序结果,用3’-Full RACE、5’-Full RACE方法获得Unigene 14872基因全长。用NCBI对其进行生物信息学分析,预测其具有丝氨酸/苏氨酸蛋白激酶(Pkinase)结构域。设计引物PCR扩增Pkinase结构域,然后克隆至pET32a(+)载体,再热转化至JM109中保存并进行质粒测序。重组质粒pET32a(+)-Pkinase构建成功,并成功诱导表达重组蛋白,初步鉴定该蛋白具有抗肿瘤活性,为进一步研究重组蛋白的活性及作用机制奠定基础。 The aim of this study is to clone the Pkinase domain in the Unigene 14872 gene of Mushroom Mushroom C91-3 transcript and perform bioinformatics analysis. Total RNA was extracted from the mycelium of Mushroom C91-3, and the full-length Unigene 14872 gene was obtained by 3’-Full RACE and 5’-Full RACE according to the transcriptome sequencing results. Bioinformatics analysis was performed with NCBI, and it was predicted to have a serine / threonine protein kinase (Pkinase) domain. Primers were designed to amplify the Pkinase domain by PCR, cloned into the pET32a (+) vector, reconstituted into JM109 and plasmid sequenced. The recombinant plasmid pET32a (+) - Pkinase was successfully constructed and the recombinant protein was successfully induced. The preliminary identification of the recombinant protein pET32a (+) - Pkin showed antitumor activity, which laid the foundation for further study on the activity and mechanism of the recombinant protein.