| 无机焦磷酸化酶基因的克隆与植物表达载体的构建 |
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| 引用本文： | 黄东杰,张树珍,冯翠莲,顾丽红,范海阔.无机焦磷酸化酶基因的克隆与植物表达载体的构建[J].热带作物学报,2007,28(2):69-73. |
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| 作者姓名： | 黄东杰 张树珍 冯翠莲 顾丽红 范海阔 |
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| 作者单位： | 中国热带农业科学院热带生物技术国家重点实验室,海口,571101 |
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| 摘 要： | 提取面包酵母基因组DNA,用无机焦磷酸化酶(PPase)基因特异引物通过PCR扩增出864bp的片段,并将该片段克隆至pMD18-T上。序列分析结果表明,该克隆序列与GeneBank上的PPase基因序列同源性达到100%。进一步将叶肉细胞特异性启动子rbcS和PPase基因克隆至植物表达载体pCBI上,构建了由rbcS启动子调控的PPase基因植物表达载体pCBIPR。通过冻融法将重组质粒导入根癌农杆菌EHA105中,为农杆菌介导的PPase基因对植物的遗传转化奠定基础。
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| 关 键 词： | 甘蔗 植物表达载体 |
| 修稿时间： | 2006-06-08 |
Cloning of Inorganic Pyrophosphatase Gene and Construction of Its Plant Expression Vector |
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| Huang Dongjie,Zhang Shuzhen,Feng Cuilian,Guo Lihong,Fan Haikuo.Cloning of Inorganic Pyrophosphatase Gene and Construction of Its Plant Expression Vector[J].Chinese Journal of Tropical Crops,2007,28(2):69-73. |
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| Authors: | Huang Dongjie Zhang Shuzhen Feng Cuilian Guo Lihong Fan Haikuo |
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| Institution: | State Key Biotechnology Laboratory for Tropical Crops, Chinese Academy of Tropical Agriculture Sciences Haikou Hainan 571101 |
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| Abstract: | An 864 bp DNA fragment was amplified by PCR using PPase gene special primer from Baker's yeast and then cloned into pMD18-T.The sequencing showed that the sequence of this fragment was 100 % homologous with that of PPase gene in GeneBank.The special promoter rbcS and PPase genes of mesophyll cells were cloned into plant expression vector pCBI,and a plant expression vector pCBIPR was constructed,in which PPase gene was controlled by the rbcS promoter.The recombination plasmids were then introduced into Agrobacterium tumefaciens EHA105 by the freezing-melting transformation method,which facilitates Agrobacterium mediated transformation of PPase gene into plant. |
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| Keywords: | Ppase rbcS |
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