牛樟芝菌丝体全长cDNA文库构建及EST序列分析  

Construction of full-length cDNA library from Antrodia cinnamomea mycelium and EST sequence analysis

作　　者：李晶[1] 陈莉 刘朋虎[1] 夏舒宁 刘艳玲 林占熺[1] LI Jing;CHEN Li;LIU Peng-hu;XIA Shu-ning;LIU Yan-ling;LIN Zhan-xi(China National Juncao Engineering Research Center,Fujian Agriculture and Forestry University,Fuzhou 350002,China;School of Life Science,Fujian Agriculture and Forestry University,Fuzhou 350002,China)

机构地区：[1]福建农林大学国家菌草工程技术研究中心,福建福州350002 [2]福建农林大学生命科学学院,福建福州350002

出　　处：《中草药》2020年第4期1052-1059,共8页Chinese Traditional and Herbal Drugs

基　　金：福建省教育厅中青年科技项目“牛樟芝深层发酵及其三萜成分差异研究”(JT180121)。

摘　　要：目的为了更好地挖掘牛樟芝Antrodiacinnamomea菌丝体三萜合成和代谢相关基因、筛选互作蛋白和牛樟芝指纹图谱分析,构建了牛樟芝菌丝体cDNA文库并分析其表达序列标签(EST)序列。方法以牛樟芝菌丝体为材料,采用Gateway法构建其cDNA文库,对部分EST序列进行生物信息学分析、功能注释和EST-SSR分析。结果成功构建了牛樟芝菌丝体cDNA文库,经鉴定文库重组率高达95%,文库滴度为6.1×106 cfu/mL,总克隆数为1.2×107 cfu,插入片段大小为300~2000 bp,平均长度达1000 bp。随机挑选单克隆进行测序获得65个有效EST序列,其中1个重叠群,64个单一序列,比对结果显示有45条序列有明确功能注释,18条为未知功能基因,GO功能注释结果表明序列涉及细胞组成、转运、催化活性、调控等功能。所有EST序列共含有271个SSR,牛樟芝核苷酸重复类型丰富,其中二核苷酸和三核苷酸重复基元占总重复基元的94.23%。结论初步明确牛樟芝菌丝体cDNA文库及其EST序列相关生物信息,为牛樟芝菌丝体基因组学研究奠定理论基础。Objective To excavate the terpenoid synthesis and metabolism-related gene function and screen the interaction protein and fingerprint analysis of Antrodia cinnamomea mycelium, a cDNA library from A. cinnamomea mycelia was constructed and the EST sequences were analyzed. Methods The cDNA library from the A. cinnamomea mycelium was constructed by the Gateway technique. A part of EST sequences about the bioinformatics, functional annotation and EST-SSR were analyzed. Results The cDNA library of the A. cinnamomea mycelium was constructed successfully. The recombinant rate of the cDNA library was 95%, the titer of the library was 6.1 × 106 cfu/mL, the total cloning number was 1.2 × 107 cfu, the length of cDNA was between 300-2 000 bp with an average length of 1 000 bp. The clones were randomly sequenced and 65 valid ESTs were obtained. After being compared in the Genbank database, 45 ESTs had a definite annotation, and 18 ESTs were unnamed and hypothetical protein. The results with GO functional annotation showed that the ESTs involved the cell composition, transport, catalytic activity, regulation functions and etc. It contained 271 SSRs of all the ESTs in total. The nucleotide repeats in A. cinnamomea were abundant, among which dinucleotide and trinucleotide repeat units were more common accounting for 94.23%. Conclusion The cDNA library from the A. cinnamomea mycelium and its ESTs related biological information were preliminarily identified, which will provide a theoretical foundation for research the mycelium genomics of A. cinnamomea.

关 键 词：牛樟芝 CDNA文库 表达序列标签 GO功能注释 SSR 

分 类 号：R282.12[医药卫生—中药学]