| 摘要: |
| [目的]研究拟南芥热激转录因子AtHsfA6a在植物细胞中的定位及其作用机制.[方法]用RT-PCR方法从野生型拟南芥总RNA中,扩增获得了849 bp的拟南芥热激转录因子AtHsfA6a cDNA片段,经过测序,结果与公布的序列完全相同.将该片段与绿色荧光蛋白(GFP)基因的cDNA重组,并克隆到表达载体pCHF3上,经PCR与酶切检测表明构建的表达载体正确.按照Clough等的方法转化拟南芥,经Kanamycin筛选和PCR检测,得到转基因植株;用激光共聚焦扫描荧光显微镜观察转基因植株幼苗的根部.[结果]在正常条件下,融合蛋白存在于细胞质中.[结论]在正常条件下拟南芥热激转录因子AtHsfA6a在细胞质中表达. |
| 关键词: 拟南芥 热激转录因子 绿色荧光蛋白 细胞定位分析 |
| DOI： |
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| 基金项目:国家重点基础研究发展规划项目（2004CB117200） |
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| Cloning and analysis of cellar location of the Arabidopsis heat transcriptional factor AtHsfA6a |
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| Abstract: |
| 【Objective】 The rsearch studied the location of the Arabidopsis heat shock transcriptional factor AtHsfA6a in the plant cell and its mechanism.【Method】 The cDNA fragment of the Arabidopsis heat shock transcriptional factor AtHsfA6a,whose length was 849 bp,was cloned by one-tube RT-PCR reaction with specific primers from Arabidopsis Columbia.Its sequence was the same as the published sequence.After the cDNA fragment was confirmed by the sequence test,it was subsequently combined with green fluorescent protein (GFP) gene,then cloned to expression vector pCHF3 transgenic Arabidopsis lines containing pCHF3-AtHsfA6a-GFP construct were generated through agrobacterium-mediated genetic transformation and obtained transgenic Arabidopsis plant through kanamycin selection and PCR test.Then the roots of transgenic Arabidopsis plant were observed through laser scanning confocal fluorescence microscop.【Result】The fusion protein lies in the cytoplasm under the ordinary condition.【Conclusion】 Under the ordinary condition,the Arabidopsis heat transcriptional factor AtHsfA6a expresses in cytoplasm of cell. |
| Key words: Arabidopsis heat shock transcriptional factor green fluorescent protein(GFP) analysis of cellar location |
