药用真菌杨树菇未知基因1C10克隆及其抗肿瘤活性初步鉴定

The Molecular Cloning and Bioactivity Characterization of Unknown Gene 1C10 from Medicinal Fungus Agrocybe aegerita

摘要: 目的：克隆杨树菇抗肿瘤相关蛋白基因，初步鉴定其活性。方法：构建cDNA文库，随机测序得到杨树菇基因1C10。高通量测序分析其在菌丝体和子实体的差异表达。构建真核表达质粒检测其在HeLa细胞中表达引起的细胞凋亡及衰老的变化，初步鉴定其抗肿瘤活性。结果：得到全长为728 bp的杨树菇基因1C10，编码107个氨基酸，高通量测序分析发现其在子实体中为高表达。在真核细胞中表达会引起肿瘤细胞的凋亡而非衰老。结论：通过构建cDNA文库及测序初步鉴定出具抗肿瘤活性的杨树菇未知基因1C10，为进一步研究其活性及机理奠定基础。 Objective: The antitumor-associated gene of Agrocybe aegerita was cloned to identify the bioactivity. Method: cDNA library was constructed and the gene 1C10 was obtained by random sequencing. The differential ex- pression of the gene was analyzed by high-throughout sequencing. The eukaryotic expression vector was constructed to detect cell apoptosis and senescence activity. Result: The gene of 1C10 was cloned with 728 bp and 107 amino acid residues. The high-throughout sequencing analyse showed that 1C10 was highly expressed in the fruiting body. The expression in HeLa cells could induce the cell apoptosis instead of senescence. Conclusion: 1C10 was identified to have antitumor activity by cDNA library and sequencing, which was fundation for further study.