一种新的用于删除选择标记基因的Cre/lox系统 |
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引用本文: | 袁媛,刘允军,王涛.一种新的用于删除选择标记基因的Cre/lox系统[J].Acta Botanica Sinica,2004,46(7):862-866. |
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作者姓名: | 袁媛 刘允军 王涛 |
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作者单位: | 中国农业大学农业生物技术国家重点实验室 北京100094
(袁媛,刘允军),中国农业大学农业生物技术国家重点实验室 北京100094(王涛) |
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基金项目: | 国家转基因植物研究与产业化专项(JY03-B-18-01),国家重点基础研究发展规划项目(2001CB1090)。~~ |
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摘 要: | 设计了一种新的诱导型Cre/lox系统,并在转基因烟草(NicotianatabacumL.)中进行了验证。在诱导剂的作用下,位于同向lox位点之间的选择标记基因(hpt)和重组酶基因(Cre)在烟草愈伤组织中被删除。在该系统中,Cre基因在玉米乙酰苯胺类化合物诱导启动子(In5-2)的控制下表达。对转基因后代的分子检测结果表明,不论是否加入了诱导剂,目的基因(gus)均被整合到烟草基因组中;在诱导剂处理的48株转基因烟草T0代中,45株的hpt基因被删除了。该系统只使用一个载体,克服了二次转化系统带来的问题。
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关 键 词: | 乙酰苯胺类化合物诱导启动子(In5-2) Cre/lox 无标记 转基因烟草 |
A New Cre/lox System for Deletion of Selectable Marker Gene |
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Authors: | YUAN Yuan LIU Yun-Jun WANG Tao |
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Institution: | YUAN Yuan,LIU Yun-Jun,WANG Tao* |
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Abstract: | A new inducible Cre/lox system was constructed in transgenic tobacco (Nicotiana tabacumL.) plants. The inducer-treatment of tobacco callus mediates an excision event in which the selectablemarker gene and Cre gene between two lox sites were deleted. A chloroacetanilide-induced promoter(In5-2) was used to control the expression of Cre gene in this system. Molecular analysis of transgenictobacco plants showed the interested gene, b-glucuronidase (gus), was integrated into the genome whetherremoving has been successful, and forty-five out of forty-eight T0 plants were transgenic tobacco withoutthe marker gene, hpt. This system uses a single vector to circumvent the flaw of other dual recombinasevector systems. |
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Keywords: | chloroacetanilide-induced promoter (In5-2) Cre/lox marker-free transgenic tobacco |
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