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.docVIP

Medical Diagnosis 医学诊断, 2012, 2, 1-5 /10.12677/md.2012.21001 Published Online March 2012 (/journal/md) The Molecular Cloning and Bioactivity Characterization of Unknown Gene 1C10 from Medicinal Fungus Agrocybe aegerita Yi Liang , Lianxian Guo , Hui Sun 1* 2* 3 1 2 3 Department of Clinical Immunology, Guangdong Medical College, Dongguan Department of Occupational and Environmental Health, Guangdong Medical College, Dongguan The College of Life Sciences, Wuhan University, Wuhan Email: liangyi@ Received: Feb. 29th, 2012; revised: Mar. 9th, 2012; accepted: Mar. 10th, 2012 Abstract: 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. Keywords: Medicinal Fungus; Antitumor Activity; cDNA Sequence 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 梁 一，郭莲仙，孙慧 3 1* 2* 1 2 3 广东医学院检验学院临床免疫教研室，东莞 广东医学院公共卫生学院劳动卫生与环境卫生教研室，东莞 武汉大学生命科学学院，武汉 Email: liangyi@ 收稿日期：2012年 2月 29日；修回日期：2012年 3月 9日；录用日期：2012年 3月 10日 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