| 利用番茄U3snRNA基因上游启动区构建植物表达载体及对烟草的转化 |
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| 引用本文： | 李丽莉,扈廷茂,扈会平,刘明秋,苏慧敏.利用番茄U3snRNA基因上游启动区构建植物表达载体及对烟草的转化[J].内蒙古大学学报(自然科学版),2005,36(1):63-67. |
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| 作者姓名： | 李丽莉 扈廷茂 扈会平 刘明秋 苏慧敏 |
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| 作者单位： | 1. 内蒙古大学生物系,呼和浩特,010021
2. 内蒙古大学生物系,呼和浩特,010021;丽珠生物工程制药厂,珠海,519000 |
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| 基金项目： | 国家自然科学基金资助项目(39760036) |
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| 摘 要： | 利用番茄U3snRNA基因上游启动区和ACC合成酶反义RNA-核酶嵌合基因DNA片段,构建含U3snRNA基因上游启动区-ACC合成酶的反义RNA-核酶嵌合序列的表达载体,重组于植物双元表达载体pGA643中,得到pGU3R.用三亲融合法导入农杆菌LBA4404中,采用叶盘法转化烟草,诱导再生小植株,获得了卡那霉素的抗性植株.提取抗性植株总DNA,通过PCR、PCRSouthern杂交检测并分别用启动区序列和ACC合成酶的反义RNA-核酶嵌合序列作探针,通过Southern杂交检测,已筛选出整合有外源基因的转化植株.为进一步研究U3snRNA上游启动区增强反义RNA-核酶基因的表达奠定了基础.
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| 关 键 词： | 烟草 U3snRNA上游启动区 反义RNA 核酶 |
| 文章编号： | 1000-1638(2005)01-0063-05 |
| 修稿时间： | 2003年9月11日 |
Construction of Plant Expression Vector with Tomato U3snRNA Promoter and Transformation of Tobacco |
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| LI Li-li,HU Ting-mao,HU Hui-ping.Construction of Plant Expression Vector with Tomato U3snRNA Promoter and Transformation of Tobacco[J].Acta Scientiarum Naturalium Universitatis Neimongol,2005,36(1):63-67. |
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| Authors: | LI Li-li HU Ting-mao HU Hui-ping |
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| Institution: | LI Li-li~1,HU Ting-mao~1,HU Hui-ping~ |
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| Abstract: | The plant expression vector pGU3R, carrying a tomato U3snRNA promoter and ACC synthase antisense RNA-Ribozyme chimeric DNA sequences, has been constructed.pGU3R was introduced into Agrobacterium tumefaciens by triparental mating. Tobacco leaf discs were transformed via Agrobacterium tumefaciens mediated procedure.The transgenic tobacco plantlets formed in induction medium and plants resistant to kanamycin were obtained.PCR and Southern blot analyses confirmed that U3snRNA promoter and ACC synthase antisense RNA-Ribozyme chimeric DNA sequence have been integrated into the genome of tobacco. |
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| Keywords: | tobacco U3sn RNA promoter antisenseRNA ribozyme |
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