过刊浏览

亚洲棉GAE6-3A上游序列的分离及其在烟草中的表达

于晓红, 朱勇清, 林芝萍, 陈晓亚, 许智宏

中科院上海植物生理研究所分子国家重点实验室.上海 200032

摘要

根据E6基因保守域设计引物,PCR扩增出亚洲棉(Gossypium arboreum L.)GAE6基因长约400bp片段,序列分析表明该片段与海岛棉(G. barbadense)E6基因同源性达96.8%。进一步合成2个反向引物协助进行PCR-96孔板筛库分离到亚洲棉GAE6-3A克隆。酶切鉴定其插入片段长约8.0kb,序列测定及分析结果表明其上游长约1.5kb。将GAE6-3A上游序列克隆至含有内含子的GUS基因前,构建了植物表达载体。三亲杂交后农杆菌介导转化烟草,组织化学分析显示GUS基因在转基因烟草植株的根、茎、叶的表皮(包括表皮毛)及维管组织表达较强。

关键词： GAE6-3A; 上游序列; 亚洲棉; 棉纤维

Isolation of GAE6-3A 5^-upstream fragment from Gossypium arboreum and its expression in tobacco

Yu Xiaohong, Zhu Yongqing, Lin Zhiping, Chen Xiaoya, Xu Zhihong

National Laboratory of Plant Molecular Genetics,Shanghai Institute of Plant Physiology,Chinese Academy of Sciences.Shanghai 200032

Abstract

Digestion with several restrictive enzymes showed that the insert is about 8.0 kb long, containing a 1.5 kb E6 5^-upstream region. Sequence alignment of G. arbireym E6 upstream fragment with upstream fragment 599 bp of an E6 gene (EMBL accession number I34396) revealed a 94.5% identity. The GAE6-3A upstream fragment was subcloned and ligated to the GUS reporter gene, then transferred into tobacco (Nicotiana tabacum) plants by Agrobacterium tumfacien mediated transformation. GUS activity was observed in the epidermis and vascular tissues of leaves, roots and stems.

Key words: GAE6-3A;Upstream fragment;Gossypium arboreum;Cotton fiber

收稿日期：　　录用日期：

通讯作者：　　E-mail:

引用本文：

于晓红, 朱勇清, 林芝萍, 陈晓亚, 许智宏. 亚洲棉GAE6-3A上游序列的分离及其在烟草中的表达[J]. 生理学报 2000; 52 (2): .

Yu Xiaohong, Zhu Yongqing, Lin Zhiping, Chen Xiaoya, Xu Zhihong. Isolation of GAE6-3A 5^-upstream fragment from Gossypium arboreum and its expression in tobacco. Acta Physiol Sin 2000; 52 (2): (in Chinese with English abstract).