水母雪莲愈伤组织cDNA文库的构建

植物学报 ›› 2004, Vol. 21 ›› Issue (01): 61-65.

水母雪莲愈伤组织cDNA文库的构建

金治平赵德修乔传令付春祥

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）

收稿日期:2002-12-29 修回日期:2003-07-07 出版日期:2004-02-20 发布日期:2004-02-20
通讯作者: 赵德修

Construction of cDNA Library from the Callus of Saussrea medusa Maxim

JIN Zhi-Ping, ZHAO De-Xiu, QIAO Chuan-Ling, FU Chun-Xiang

1(Institute of Botany, the Chinese Academy of Sciences, Beijing 100093)2(Institute of Zoology, the Chinese Academy of Sciences, Beijing 100090)

Received:2002-12-29 Revised:2003-07-07 Online:2004-02-20 Published:2004-02-20
Contact: ZHAO De-Xiu

摘要/Abstract

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。

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.

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. 植物学报, 2004, 21(01): 61-65.

JIN Zhi-Ping; ZHAO De-Xiu; QIAO Chuan-Ling; FU Chun-Xiang. Construction of cDNA Library from the Callus of Saussrea medusa Maxim. Chinese Bulletin of Botany, 2004, 21(01): 61-65.

导出引用管理器 EndNote|Ris|BibTeX

链接本文: https://www.chinbullbotany.com/CN/

https://www.chinbullbotany.com/CN/Y2004/V21/I01/61