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在转基因烟草中表达功能单叶植物光敏色素。

Expression of a functional monocotyledonous phytochrome in transgenic tobacco.

机构信息

Agricultural Products Department, E.I. du Pont de Nemours & Co., PO Box 80402, Wilmington, DE 19880-0402.

出版信息

EMBO J. 1989 Apr;8(4):1005-12. doi: 10.1002/j.1460-2075.1989.tb03467.x.

Abstract

A chimeric oat phytochrome structural gene with an uninterrupted coding region was constructed for expression of the monocot protein in transgenic plants. The structural gene was placed under the transcriptional control of either a light-regulated oat phytochrome promoter or the constitutively active cauliflower mosaic virus 35S promoter. These genes were then introduced into Nicotiana tabacum and N.plumbaginifolia. None of the regenerated plants showed expression of oat phytochrome RNA when transcription was controlled by the oat promoter. In contrast, RNA was obtained in plants when the structural gene was functionally linked to the 35S promoter. Transformants expressing oat phytochrome RNA produced a full length 124-kd polypeptide that was recognized by oat-specific anti-phytochrome monoclonal antibodies. The oat protein was a substrate for chromophore addition in tobacco as judged by its red/far-red photoreversible sensitivity to trypsin degradation. Production of oat phytochrome in transgenic plants gave rise to increased phytochrome spectral activity in both light- and dark-grown plants. This increased phytochrome content resulted in phenotypic changes in transformed plants, including semi-dwarfism, darker green leaves, increased tillering and reduced apical dominance. The possible significance of expressing a biologically active phytochrome in transgenic plants is discussed.

摘要

为了在转基因植物中表达单子叶植物蛋白,构建了一个具有完整编码区的嵌合燕麦光敏色素结构基因。该结构基因受光调控的燕麦光敏色素启动子或组成型活性的花椰菜花叶病毒 35S 启动子的转录调控。然后将这些基因导入烟草和烟草(N.plumbaginifolia)。当转录受燕麦启动子控制时,没有再生植物表现出燕麦光敏色素 RNA 的表达。相比之下,当结构基因与 35S 启动子功能连接时,在植物中获得了 RNA。表达燕麦光敏色素 RNA 的转化体产生了全长 124kDa 的多肽,该多肽被燕麦特异性抗光敏素单克隆抗体识别。根据其对胰蛋白酶降解的红/远红光可逆敏感性,判断烟草中的燕麦蛋白是发色团添加的底物。在转基因植物中产生燕麦光敏素导致光和暗培养的植物中光敏素光谱活性增加。这种增加的光敏素含量导致转化植物发生表型变化,包括矮化、叶片颜色更深、分蘖增加和顶端优势降低。讨论了在转基因植物中表达具有生物活性的光敏素的可能意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/243d/400907/7e8ac8743f73/emboj00128-0019-a.jpg

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