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转基因烟草中热诱导潮霉素抗性

Heat-inducible hygromycin resistance in transgenic tobacco.

作者信息

Severin K, Schöffl F

机构信息

Lehrstuhl für Genetik, Fakultät für Biologie, Universität Bielefeld, FRG.

出版信息

Plant Mol Biol. 1990 Dec;15(6):827-33. doi: 10.1007/BF00039423.

Abstract

We have constructed a chimaeric gene consisting of the promoter of the soybean heat shock (hs) gene Gmhsp17, 6-L, the coding region of a hygromycin phosphotransferase (hpt) gene, and the termination sequence of the nopaline synthase (nos) gene. This gene fusion was introduced into tobacco by Agrobacterium-mediated gene transfer. Heat-inducible synthesis of mRNA was shown by northern hybridization, and translation of this RNA into a functional protein was indicated by plant growth on hygromycin-containing media in a temperature-dependent fashion. One hour incubation at 40 degrees C per day, applied for several weeks, was sufficient to express the resistant phenotype in transgenic plants containing the chimaeric hs-hpt gene. These data suggest that the hygromycin resistance gene is functional and faithfully controlled by the soybean hs promoter. The suitability of these transgenic plants for selection of mutations that alter the hs response is discussed.

摘要

我们构建了一个嵌合基因,它由大豆热激(hs)基因Gmhsp17,6-L的启动子、潮霉素磷酸转移酶(hpt)基因的编码区以及胭脂碱合酶(nos)基因的终止序列组成。通过农杆菌介导的基因转移将这个基因融合体导入烟草。Northern杂交显示了热诱导的mRNA合成,并且这种RNA在含潮霉素的培养基上以温度依赖的方式转化为功能性蛋白质,从而表明了植物的生长情况。每天在40摄氏度下培养1小时,持续数周,足以在含有嵌合hs-hpt基因的转基因植物中表达抗性表型。这些数据表明潮霉素抗性基因是有功能的,并且受大豆hs启动子的忠实调控。讨论了这些转基因植物对于选择改变热激反应的突变体的适用性。

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