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利用烟草病程相关蛋白基因启动子区域在转基因莴苣植物中诱导嵌合基因构建体的表达。

Induced expression of a chimeric gene construct in transgenic lettuce plants using tobacco pathogenesis-related protein gene promoter region.

机构信息

National Institute of Agrobiological Resources, Kannondai, Tsukuba, 305, Ibaraki, Japan.

出版信息

Plant Cell Rep. 1990 Jun;9(1):6-9. doi: 10.1007/BF00232124.

Abstract

The expression of a stress- and salicylic acidinducible protein gene from tobacco, PR1a protein gene, was determined after its Introduction to lettuce (Lactuca sativa L.) plants. The 5' flanking 2.4 Kb fragment from PR1a gene was joined to the bacterial β -glucuronidase (GUS) gene (PR-GUS) and introduced into lettuce cotyledons by Agrobacterium-mediated gene transfer using a binary vector containing a kanamycin-resistance gene as a selectable marker. As a control with constitutive expression, the chimeric gene consisting of CaMV 35S RNA promoter and GUS gene (35S-GUS) was used. An improved method for shoot formation directly from lettuce cotyledons was used effectively for transformation, shortening the time for regeneration. In 70% or more of kanamycin-resistant regenerated lettuce plants, into which PR-GUS or 35S-GUS was introduced, high GUS activity and integration of the chimeric gene into the lettuce genome were detected. By treatment with salicylic acid, GUS activity increased 3- to 50-fold in PR-GUS transformants, however, no increase was detected in 35S-GUS plants. These results showed that the promoter of the stress-inducible tobacco PR1a protein gene was introduced into lettuce plants, and the introduced chimeric gene was expressed normally under the regulated control of the PRla promoter.

摘要

将烟草中一种受应激和水杨酸诱导的蛋白基因 PR1a 蛋白基因的表达导入生菜(Lactuca sativa L.)植物后进行了测定。将 PR1a 基因的 5'侧翼 2.4 Kb 片段与细菌β-葡萄糖醛酸酶(GUS)基因(PR-GUS)连接,并通过含有卡那霉素抗性基因作为选择标记的二元载体,利用农杆菌介导的基因转移将其导入生菜子叶。作为组成型表达的对照,使用了由 CaMV 35S RNA 启动子和 GUS 基因(35S-GUS)组成的嵌合基因。一种改良的直接从生菜子叶形成芽的方法有效地用于转化,缩短了再生时间。在 70%或更多的卡那霉素抗性再生生菜植物中,导入了 PR-GUS 或 35S-GUS,检测到高 GUS 活性和嵌合基因整合到生菜基因组中。用水杨酸处理后,PR-GUS 转化体中的 GUS 活性增加了 3 到 50 倍,但在 35S-GUS 植物中没有检测到增加。这些结果表明,应激诱导的烟草 PR1a 蛋白基因的启动子已被导入生菜植物,并且在 PRla 启动子的调控控制下,引入的嵌合基因正常表达。

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