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带内含子卡那霉素抗性基因双元载体构建及烟草转化

彭日荷, 黄晓敏, 李贤, 孙爱君, 姚泉洪, 彭友良

上海农业科学院生物技术中心,上海市农业遗传重点实验室.上海 201106；中国农业大学

摘要

农杆菌介导法是植物基因转化的常用方法,然而由于筛选培养基中常用的抗生素头孢霉素和羧苄青霉素具有类植物激素活性,影响外植体的再生和转化频率。将一个植物的内含子的卡那霉素抗性基因。构建带该基因的植物元双表达载体pYP1202并转化烟草,受侵外植体在含卡那霉素50-200mg/L的选择培养基中抗性芽分化频率不受卡那霉素浓度影响,然而具有GUS活性的转化子占分化芽的比例却随着卡那霉素浓度的增加而升高。当培养基中加入500mg/L羧苄青霉素后受侵外植体产生的抗性芽频率比单一的卡那霉素筛选提高近1倍,高达91.4%,然而具GUS活性的转化子占抗性芽的比例仅有26.7%,在200mg/L的卡那霉素筛选下,比例升至93.3%。用带内含子卡那霉素抗性基因构建的植物表达载体转化植物可以减少假抗性芽的产生。

关键词： 带内含子卡那霉素抗性基因; GUS活性; 烟草转化

Construction of a plant binary expression vector containing intron-kanamycin gene and transformation in Nicotiana tabacum

Peng Rihe, Huang Xiaomin, Li Xian, Sun Aijun, Yao Quanhong, Peng Youliang

Shanghai Key Laboratory of Agricultural Genetic and Breeding,Agro-Biotechnology Research Center,Shanghai Academy of Agricultural Sciences.Shanghai 201106

Abstract

From these results, it can be concluded that the intron-kanamycin gene can be used as an appropriated gene in Agrobacterium-mediated plant transformation.

Key words: Intron-kanamycin gene;GUS activity;Transformation in tobacco

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引用本文：

彭日荷, 黄晓敏, 李贤, 孙爱君, 姚泉洪, 彭友良. 带内含子卡那霉素抗性基因双元载体构建及烟草转化[J]. 生理学报 2001; 53 (1): .

Peng Rihe, Huang Xiaomin, Li Xian, Sun Aijun, Yao Quanhong, Peng Youliang. Construction of a plant binary expression vector containing intron-kanamycin gene and transformation in Nicotiana tabacum. Acta Physiol Sin 2001; 53 (1): (in Chinese with English abstract).