| OsMAPK4基因的克隆、序列分析及其植物表达载体的构建 |
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| 引用本文： | 李杰,朱延明,齐岩,代翠红,柏锡.OsMAPK4基因的克隆、序列分析及其植物表达载体的构建[J].东北农业大学学报,2005,36(3):324-328. |
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| 作者姓名： | 李杰 朱延明 齐岩 代翠红 柏锡 |
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| 作者单位： | 东北农业大学生命科学学院,黑龙江,哈尔滨,150030;东北农业大学生命科学学院,黑龙江,哈尔滨,150030;东北农业大学生命科学学院,黑龙江,哈尔滨,150030;东北农业大学生命科学学院,黑龙江,哈尔滨,150030;东北农业大学生命科学学院,黑龙江,哈尔滨,150030 |
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| 基金项目： | 国家重点基础研究发展计划(973计划)，国家自然科学基金，黑龙江省博士后科研启动基金 |
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| 摘 要： | 以低温处理的耐盐水稻辽盐241植株叶片总RNA为模板,用OsMAPK4基因特异引物通过RT-PCR扩增出1500bp的片段,并将该片段克隆至pUC18上。序列分析结果表明,该克隆序列与GenBank上的OsMAPK4基因序列同源性达99.4%,氨基酸同源性达99.1%。进一步将OsMAPK4基因分别克隆至植物表达载体卡盒pBCE12和pBC29A上,构建了分别由E12启动子、rd29A启动子调控的OsMAPK4基因植物表达载体pBME12和pBM29A。通过冻融法将重组质粒导入根癌农杆菌LBA4404中,为农杆菌介导法转化植物,分析OsMAPK4基因的功能奠定了基础。
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| 关 键 词： | OsMAPK4基因 基因克隆 植物表达载体 |
| 文章编号： | 1005-9369(2005)03-0324-05 |
| 修稿时间： | 2004年7月5日 |
Cloning and sequencing of OsMAPK4 gene and construction of its plant expression vector |
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| LI Jie,ZHU Yan-ming,QI Yan,DAI Cui-hong,BAI Xi.Cloning and sequencing of OsMAPK4 gene and construction of its plant expression vector[J].Journal of Northeast Agricultural University,2005,36(3):324-328. |
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| Authors: | LI Jie ZHU Yan-ming QI Yan DAI Cui-hong BAI Xi |
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| Abstract: | A 1 500 bp DNA fragment amplified by RT-PCR use OsMAPK4 gene special primer from liaoyan241 leaf treated by low temperature was cloned into pUC18. The result of sequencing showed that the sequence of this fragment was 99.4% homologous with that of OsMAPK4 gene in GenBank and amino acid homologous is 99.1%. Further more OsMAPK4 gene was cloned into plant express vector cassette pBCE12 and pBC29A. Thus two plant express vectors, pBME12 and pBM29A were constructed, in which OsMAPK4 gene was controlled by E12 promoter and rd29A promoter respectively. Then the recombination plasmids were introduced into Agrobacterium LBA4404 by freezing-melting transformation method. This work provides a foundation for transferring OsMAPK4 gene into plant by Agrobacterium-mediated method and its function analysis. |
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| Keywords: | OsMAPK4 gene gene cloning plant express vector |
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