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生长素结合蛋白cDNA的克隆及其在烟草中的表达

张洪霞, 白永延, 许智宏

中科院上海植物生理研究所,植物分子遗传国家重点实验室.上海 200032

摘要

基于拟南芥内质网生长素结合蛋白基因的cDNA序列,设计合成了Ap5和Ap3两个引物,应用 RT-PCR 技术扩增了拟南芥的ABP基因。将该基因克隆在植物表达载体 p35SSIN 的35 S启动子和 Nos 3^端之间,得到植物表达载体 p35SE。通过农杆菌介导的方法对烟草SR1进行了转化,由分子杂交等检测证明,生长素结合蛋白基因已在烟草中表达,同时转基因烟草后代对生长素的敏感性明显增加。

关键词： 生长素结合蛋白; 反转录-聚合酶链式反应; 转基因植株

Cloning of auxin binding protein cDNA and its expression in tobacco

Zhang Hongxia, Bai Yongyan, Xu Zhihong

Shanghai Institute of Plant Physiology, The Chinese Academy of Sciences. Shanghai 200032

Abstract

According to the reported coding sequence of auxin binding protein(ABP) located in the endoplasmic reticulum of Arabidopsis, we have designed and synthesized two primers Ap5 and Ap3. The whole sequence of ABP cDNA was cloned by RT-PCR and confirmed by DNA sequencing.Plant expression vector p35SE was constructed to transform the explants of tobacco(SR1) leaves by Agrobacterium-mediated method.Southern and Northern analysis confirmed that the foreign ABP gene has been expressed in tobacco plant cells. It was also found that the progenies of p35SE transgenic tobacco plants were significantly auxin-sensitive.

Key words: Auxin binding protein;RT-plymerase chain reaction;Transgenic plant

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引用本文：

张洪霞, 白永延, 许智宏. 生长素结合蛋白cDNA的克隆及其在烟草中的表达[J]. 生理学报 1997; 49 (2): .

Zhang Hongxia, Bai Yongyan, Xu Zhihong. Cloning of auxin binding protein cDNA and its expression in tobacco. Acta Physiol Sin 1997; 49 (2): (in Chinese with English abstract).