药学学报, 2010, 45(4): 524-529
引用本文:
李 滢 孙 超 罗红梅 李西文 牛云云 陈士林. 基于高通量测序454 GS FLX的丹参转录组学研究[J]. 药学学报, 2010, 45(4): 524-529.
LI Ying, Sun- Chao, Luo-Gong-Mei, Li-Xi-Wen, Niu-Yun-Yun, Chen-Shi-Lin. Transcriptome characterization for Salvia miltiorrhiza using 454 GS FLX[J]. Acta Pharmaceutica Sinica, 2010, 45(4): 524-529.

基于高通量测序454 GS FLX的丹参转录组学研究
李 滢1, 孙 超1, 罗红梅1, 李西文1, 2, 牛云云1, 陈士林1*
(1. 中国医学科学院、北京协和医学院药用植物研究所, 北京 100193; 2. 清华大学化学系, 北京 100084)
摘要:

本研究应用新一代高通量测序技术454 GS FLX Titanium2年生丹参根的转录组进行测序, 研究其基因表达谱, 挖掘其功能基因。获得46 722表达序列标签 (express sequence tags, EST), 序列平均长度414 bp, Sanger测序的长度相当。所得序列与GenBank丹参EST合并拼接, 获得18 235unigene, 其中, 454高通量测序发现了13 980条新的unigene。数据库中的序列同源性比较表明, 其中73.0% (13 308) 与其他生物的已知基因具有不同程度的同源性。通过BLASTGene Ontology分析获得了可能参与丹参酮合成的序列27 (编码15个关键酶), 参与丹酚酸合成的序列29 (编码11个关键酶), 细胞色素P450序列70, 转录因子序列577条。454高通量测序技术作为药用植物功能基因组研究的重要手段可在丹参功能基因的发现中发挥重要作用, 这些基因的发现为丹参酮和丹酚酸类化合物生物合成研究奠定了基础, 同时也为丹参的转录组研究提供了基础数据。

关键词:   
Transcriptome characterization for Salvia miltiorrhiza using 454 GS FLX
Abstract:

To investigate the profile of gene expression in Salvia miltiorrhiza and elucidate its functional gene, 454 GS FLX platform and Titanium regent were used to produce a substantial expressed sequence tags (ESTs) dataset from the root of S. miltiorrhiza.  A total of 46 722 ESTs with an average read length of 414 bp were generated.  454 ESTs were combined with the S. miltiorrhiza ESTs from GenBank.  These ESTs were  assembled into 18 235 unigenes.  Of these unigenes, 454 sequencing identified 13 980 novel unigenes.  73% of these unigenes (13 308) were annotated using BLAST searches (E-value 1e-5) against the SwissProt, KEGG, TAIR, Nr and Nt databases.  Twenty-seven unigenes (encoding 15 enzymes) were found to be involved in   tanshinones biosynthesis, and 29 unigenes (encoding 11 enzymes) involved in phenolic acids biosynthesis.   Seventy putative genes were found to encode cytochromes P450 and 577 putative transcription factor genes.  Data presented in this study will constitute an important resource for the scientific community that is interested in the molecular genetics and functional genomics of S. miltiorrhiza.

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