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玉米泛素基因启动子-bar 嵌合基因在转基因水稻植株中的表达。

Expression of a Maize Ubiquitin Gene Promoter-bar Chimeric Gene in Transgenic Rice Plants.

机构信息

Department of Botany, Faculty of Science, Hokkaido University, Sapporo 060, Japan.

出版信息

Plant Physiol. 1992 Nov;100(3):1503-7. doi: 10.1104/pp.100.3.1503.

Abstract

We have constructed a chimeric gene consisting of the promoter, first exon, and first intron of a maize ubiquitin gene (Ubi-1) and the coding sequence of the bar gene from Streptomyces hygroscopicus. This construct was transferred into rice (Oryza sativa L.) protoplasts via electroporation, and 10 plants were regenerated from calli that had been selected for resistance to exogenously supplied bialaphos. Transgenic plants grown in a greenhouse were resistant to both bialaphos and phosphinothricine at a dosage lethal to untransformed control plants. Evidence of stable integration of the transferred gene into the genome of the regenerated primary transformant plants was obtained from Southern blot analysis. In addition, northern blot analysis indicated expression and proper splicing of the maize ubiquitin gene first intron from the primary chimeric transcript in these transgenic rice plants, and western blot analysis and enzymic assays verified expression of the active bar gene product. Apparent mendelian segregation for bialaphos resistance in T(1) progeny of primary transformants was confirmed.

摘要

我们构建了一个嵌合基因,该基因由玉米泛素基因(Ubi-1)的启动子、第一外显子和第一内含子以及吸水链霉菌的 bar 基因的编码序列组成。该构建体通过电穿孔转染到水稻(Oryza sativa L.)原生质体中,并从对外源供应的草丁膦抗性选择的愈伤组织中再生了 10 株植物。在温室中生长的转基因植物对草丁膦和膦丝菌素都具有抗性,而剂量对未转化的对照植物具有致死作用。从 Southern blot 分析中获得了整合到再生初级转化体植物基因组中的转移基因的稳定证据。此外, northern blot 分析表明,在这些转基因水稻植物中,从初级嵌合转录本中表达和正确剪接了玉米泛素基因的第一内含子,western blot 分析和酶测定验证了活性 bar 基因产物的表达。初级转化体 T(1)代对草丁膦抗性的明显孟德尔分离得到了确认。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/48de/1075812/fa76c2a423b2/plntphys00711-0431-a.jpg

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