| 水母雪莲愈伤组织cDNA文库的构建 |
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| 引用本文： | 金治平,赵德修,乔传令,付春祥.水母雪莲愈伤组织cDNA文库的构建[J].植物学报,2004,21(1):61-65. |
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| 作者姓名： | 金治平 赵德修 乔传令 付春祥 |
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| 作者单位： | 1（中国科学院植物研究所 北京 100093) 2 (中国科学院动物研究所 北京 100090） |
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| 基金项目： | 国家自然科学基金,中国科学院知识创新工程项目 |
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| 摘 要： | 用TRIZOL Reagent提取水母雪莲红色系1~15 d愈伤组织总RNA，用SMART cDNA Library Construction Kit构建cDNA文库。经测定原始文库滴度达到1.5~4×106，扩增总文库滴度达到1011，重组率达到98%，插入片段在0.5 kb到3 kb之间，多在1 kb左右。通过PCR检测，从总文库中检测到了雪莲CHS、DFR及SmP基因的特异片段。SmP基因是转录调控因子，其表达丰度很低。各项指标都表明，已获得高质量的cDNA文库，为雪莲基因资源保存，雪莲类黄酮次生代谢分子调控奠定了坚实的基础。
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| 关 键 词： | 水母雪莲 愈伤组织 cDNA文库 |
| 收稿时间： | 2002-12-29 |
| 修稿时间： | 2003-07-07 |
Construction of cDNA Library from the Callus of Saussrea medusa Maxim |
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| JIN Zhi-Ping,ZHAO De-Xiu,QIAO Chuan-Ling,FU Chun-Xiang.Construction of cDNA Library from the Callus of Saussrea medusa Maxim[J].Bulletin of Botany,2004,21(1):61-65. |
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| Authors: | JIN Zhi-Ping ZHAO De-Xiu QIAO Chuan-Ling FU Chun-Xiang |
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| Institution: | 1(Institute of Botany, the Chinese Academy of Sciences, Beijing 100093)2(Institute of Zoology, the Chinese Academy of Sciences, Beijing 100090) |
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| Abstract: | Total RNA of 1~15 d red line callus of S.medusa was extracted by TRIZOL Reagent. cDNA library was constructed with SMART cDNA Library Construction Kit. The primary library has a high titer from 1.5×106 to 4×106, in which 98% clones are recombinant and the insert cDNAs are from 0.5 kb to 3 kb. The amplified library has a titer of 1011. The positive signals of CHS, DFR and SmP gene were detected by PCR in the amplified library, though SmP gene was expressed at low abundance . This high quality cDNA library provides a useful tool for the study of the molecular mechanisms of the secondary metabolism of the flavonoids of S.medusa. |
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| Keywords: | , |
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