中国农业科学2011，Vol.44Issue(17)：3491-3500,10.DOI:10.3864/j.issn.0578-1752.2011.17.001
基于AlcR/alcA和Cre/loxP系统的标记基因诱导删除体系
Excision of Selectable Markers Based on Inducible AlcR/alcA and Cre/loxP Systems
摘要
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.关键词
Cre/loxP重组系统/AlcR/alcA诱导系统/删除/选择标记基因Key words
Cre/loxP recombination system/ AlcR/alcA induce system/ excision/ selectable markers引用本文复制引用
赵青,郭仰东,谢华,马荣才,姚磊..基于AlcR/alcA和Cre/loxP系统的标记基因诱导删除体系[J].中国农业科学,2011,44(17):3491-3500,10.基金项目
国家“863”计划项目 (2008AA10Z154,2009AA10Z104-2)、北京市科技计划(Z07070501770704)、北京市农林科学院科技创新能力建设专项(KJCX201102003)、中央高校基本科研业务费专项资金(2009-2-06) （2008AA10Z154,2009AA10Z104-2）
