›› 2010, Vol. 12 ›› Issue (6): 109-113.DOI: 10.3969/j.issn.1008-0864.2010.06.19

• 研究报告 • 上一篇    下一篇

转基因香石竹Moonlite品系特异性定性PCR检测方法的建立

李鹏*,潘爱虎*,贾军伟,蒋玲曦,白蓝,唐雪明   

  1. (上海市农业科学院生物技术研究所, 上海市农业遗传育种重点实验室, 农业部转基因植物环境安全监督检验测试中心(上海), 上海 201106)
  • 收稿日期:2010-08-09 修回日期:2010-08-23 出版日期:2010-12-15 发布日期:2010-10-14
  • 通讯作者: 唐雪明,研究员,Tel:021-62208750; E-mail:xueming70@gmail.com
  • 作者简介:*本文共同第一作者。李鹏,研究实习员,硕士,研究方向为转基因植物环境安全性评价。E-mail:sunsite@126.com。潘爱虎,副研究员,研究方向为转基因成分检测及转基因植物环境安全评价。Tel:021-62208750|E-mail:aihup@yahoo.com.cn。
  • 基金资助:

    上海市科技兴农重点攻关项目\[沪农科攻字(2008)8-9号,沪农科攻字(2009)6-4-2号\];上海市科委创新平台建设项目(10DZ2294103);上海市农科院青年科技发展基金\[农青年科技2010(15)\]资助。

Establishment of Event-Specific Qualitative PCR for Detecting Transgenic Carnation ‘Moonlite’

LI Peng*, PAN Ai-hu*, JIA Jun-wei, JIANG Ling-xi, BAI Lan, TANG Xue-ming   

  1. (Biotech Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai Key Laboratory of Agricultural Genetics and Breeding, Supervision, Inspection and Test Center for Environmental Safety of GM Crops(Shanghai), Ministry of Agriculture, Shanghai 201106, China)
  • Received:2010-08-09 Revised:2010-08-23 Online:2010-12-15 Published:2010-10-14

摘要:

以转基因香石竹品种Moonlite(月之伊人)为研究对象,通过TAIL-PCR方法,获得外源基因插入位点的左侧旁邻序列。根据旁邻序列设计引物,建立品系特异性定性PCR方法,其扩增片段覆盖了转化载体及香石竹基因组临界序列。本方法特异性好、灵敏度高,可快速、准确、高效地检测转基因香石竹Moonlite品种。

关键词: 转基因;香石竹;品系特异性;PCR检测

Abstract:

An event-specific transgenic detection method for ‘Moonlite’ was established. By thermal asymmetric interlaced PCR (TAIL-PCR) technique, the flanking sequences of the exogenous integration in the genome of ‘Moonlite’ were cloned and sequenced with the specific nested primers based on the left border. According to the left flanking sequence, the event-specific primers were designed to amplify the fragments, which could span the exogenous DNA and carnation genome and the event-specific qualitative PCR detection method for transgenic carnation ‘Moonlite’ was established. The results showed that the qualitative PCR assay was accurate, rapid and efficient for detection of ‘Moonlite’.

Key words: transgenic, carnation, event-specific, PCR detection

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