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Acta Agron Sin ›› 2010, Vol. 36 ›› Issue (2): 361-364.doi: 10.3724/SP.J.1006.2010.00361

• RESEARCH ACTIVITIES • Previous Articles    

Analysis of Junction Sequence in the Transgenic Maize MON8817 and the Methods of Qualitative PCR Detction

YUAN Lei1,2,SUN Hong-Wei1,ZHAO Lei2,YANG Chong-Liang1,SHANG You-Fen1,LU Xing-Bo1*   

  1. 1Institute of Plant Protection，Shandong Academy of Agricultural Sciences，Jinan 250100，China;2College of Life Science,Shandong Normal University,Jinan 250014,China
  • Received:2009-07-06 Revised:2009-09-06 Online:2010-02-10 Published:2009-11-17
  • Contact: LU Xing-Bao, E-mail: luxb99@sina.com, Tel: 0531-83179095

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Abstract:

The experiment was conducted to investigate the integration site of transgene in maize MON88017 and to establish event specific methods for qualitative detection of MON88017 based on the left border junction fragment, which was isolated with the amended GenomeWalker and Nested-PCR methods. Sequence alignment between the T-DNA sequence and isolated junction fragments showed a 504 bp junction fragment of MON88017 including 336 bp of T-DNA sequence and 168 bp of MON88017 genome DNA. MON88017 event-specific qualitative PCR method was established with the primers (MON88017-1F/R) targeting the junction regions to produce a 446 bp product. The limit of detection for qualitative PCR assay was 0.1%. The method developed in this work is highly specific, sensitive and suitable for MON88017 sample detection.

Key words: GMO, Event-specific detection, Qualitative PCR, MON88017 event, Left border junction fragments

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