用于植物基因研究的mCherry表达载体构建及定位表达

发布时间：2018-05-16 13:29

  本文选题：mCherry + 红色荧光蛋白　； 参考：《植物研究》2017年02期

【摘要】：荧光蛋白在生物学研究中具有广泛的应用和重要的作用,其中红色荧光蛋白mCherry因其颜色和良好的特性,对于植物基因研究具有重要的使用价值,本研究将mCherry基因构建到pBI121植物表达载体系统中,构建了pBI121MCS-mCherry载体。利用基因枪转化法转入洋葱表皮进行表达验证,显微镜观察结果显示整个洋葱细胞具有红色荧光,证明该载体能够在植物细胞中表达红色荧光蛋白。利用双酶切连接法将转录因子BpMYB4基因构建到该载体上,得到融合表达载体pBI121MCS-mCherry-BpMYB4,在洋葱表皮中表达,结果显示细胞核具有红色荧光,证明该载体能够准确表达融合蛋白,进行亚细胞定位。同时融合基因时不再需要中间载体,构建简便,引入的KpnⅠ酶切位点,增加了可选择性。因此该载体可用于植物基因表达定位及转基因植株筛选研究中,为今后的白桦基因组学研究提供了材料。
[Abstract]:Fluorescent protein (mCherry) has been widely used and played an important role in biological research. Red fluorescent protein (mCherry) has important application value for plant gene research because of its color and good characteristics. In this study, mCherry gene was constructed into pBI121 plant expression vector system and pBI121MCS-mCherry vector was constructed. The results of microscopical observation showed that the whole onion cells had red fluorescence, which proved that the vector could express red fluorescent protein in plant cells. The transcription factor BpMYB4 gene was constructed into the vector by double enzyme digestion, and the fusion expression vector pBI121MCS-mCherry-BpMYB4 was obtained, which was expressed in onion epidermis. The results showed that the cell nucleus had red fluorescence, which proved that the vector could express the fusion protein accurately. Subcellular localization was performed. At the same time, the intermediate vector is no longer needed in the fusion gene, so it is easy to construct, and the Kpn 鈪，

本文编号：1896984