Scientia Agricultura Sinica ›› 2011, Vol. 44 ›› Issue (17): 3491-3500.doi: 10.3864/j.issn.0578-1752.2011.17.001

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS •     Next Articles

Excision of Selectable Markers Based on Inducible AlcR/alcA and Cre/loxP Systems

ZHAO  Qing, GUO  Yang-Dong, XIE  Hua, MA  Rong-Cai, YAO  Lei   

  1. 1. 中国农业大学农学与生物技术学院
    2. 北京市农林科学院农业生物技术研究中心
  • Received:2011-01-07 Revised:2011-02-21 Online:2011-09-01 Published:2011-03-16
  • Contact: lei yao E-mail:yaolei@baafs.net.cn

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Abstract: 【Objective】To eliminate the potential risk in safety raised by selectable markers, an ethanol inducible excision system of selectable markers was constructed, which can be used during plant growth and development. 【Method】 The selectable markers can be removed based on AlcR/alcA inducible system and Cre/loxP site-specific recombination system. In the presence of the exogenous inducer, the activation of the downstream Cre gene was enabled. Cre recombinase identified and catalyzed excision of the intervening sequence between two directly oriented loxP sites, including selectable marker gene, AlcR/alcA and Cre/loxP system. The gene of interest, gus, was constitutively expressed before and after induction. 【Result】 The Arabidopsis thaliana transgenic plants were induced by ethanol. The results revealed that ethanol tightly control the “on” and “off” of the expression of Cre gene. After induction, the transgenic plants could not continuously grow on selective medium, which indicate the selectable marker was removed efficiently. The molecule analysis revealed the DNA fragment between two directly oriented loxP sites has been excised. 【Conclusion】 The results demonstrate that the excision of selectable markers based on inducible AlcR/alcA and Cre/loxP systems is reliable, and has a bright future.

Key words:

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