| 摘要: |
| 【目的】从玉米幼根基因组DNA中克隆β-葡萄糖苷酶基因根部特异性启动子序列ZmGLU1P，并对其功能进行分析。【方法】利用PCR技术从玉米品种 P138 幼根基因组DNA中克隆玉米根部特异性启动子片段ZmGLU1P，将其与GUS基因融合，构建植物表达载体pCAMBIA121-ZmGLU1P，转化到 EHA105根癌农杆菌中，通过根癌农杆菌介导法转化烟草NC89，对转化烟草植株进行PCR和Southern杂交检测。采集PCR和Southern杂交检测为阳性的转基因烟草的根、茎、叶，进行GUS活性的组织染色检测。【结果】克隆获得了ZmGLU1P片段，其长度为1 846 bp，与已报道的序列同源性达99%以上。转基因烟草植株的PCR 和Southern杂交结果显示,成功地获得了转基因阳性植株；GUS活性检测表明，根中GUS活性最强，而在茎和叶等组织中GUS活性甚微，表明ZmGLU1P片段具有根部特异性启动子功能。【结论】玉米β-葡萄糖苷酶基因上游1 846 bp 的片段ZmGLU1P具有根部特异性启动子功能，为根部特异性启动子。 |
| 关键词: 玉米；根部特异性启动子；功能分析；GUS染色 烟草 |
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| 基金项目:国家转基因专项(20082X08003-005)；吉林省财政厅项目(200806)；吉林省科技厅成果转化补助项目(20095044) |
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| Cloning and functional identification of the root specific promoter from corn |
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| Abstract: |
| 【Objective】 This paper aimed to clone and identify the root-specific promoter of ZmGLU1P gene from corn.【Method】 The promoter region of root-specific gene was isolated from the genomic DNA of corn P138 by PCR method,and obtained the promoter fragment ZmGLU1P.The cloned promoter was fused to the GUS reporter gene to construct plant expression vector pCAMBIA121-ZmGLU1P,which was transferred into tobacco(Nicotiana tabacum) NC89 by Agrobacteriumtum efaciens mediated method and several transformed plants were obtained.Then they were detected by PCR and Southern blotting.The root,stem,and leaf of gram positive recombinant corns were performed with GUS activity assay.【Result】The results showed that the length of the clone was 1 846 bp,and the sequence was more than 99% homology compared with the reported promoters.PCR and Southern blot results showed that the transgenic plants were obtained successfully.GUS activity assays of different organs indicated that the expression of GUS was active mostly in roots,suggesting that the ZmGLU1P gene is root-specific promoter.【Conclusion】 The cloned 1846bp fragment β-glucosidase gene from corn is root-specific promoter. |
| Key words: corn root specific promoter functional identification GUS staining tobacoo |
