萝卜水通道蛋白基因RsAQP1b的表达模式及功能分析

发布时间：2018-02-20 22:08

本文关键词： 萝卜水通道蛋白表达模式过表达　出处：《中国蔬菜》2017年05期 　论文类型：期刊论文

【摘要】：以萝卜品种白玉春为材料,利用同源克隆的方法获得萝卜水通道蛋白基因Rs AQP1b的编码序列全长,在萝卜苗期干旱胁迫下分析Rs AQP1b基因及蛋白的表达模式;在烟草中表达该基因,研究干旱胁迫下烟草中MDA含量及保护酶的活性变化。结果表明:萝卜水通道蛋白基因Rs AQP1b的编码序列全长861bp,编码286个氨基酸。制备了Rs AQP1b多克隆抗体,效价在1∶512000以上。RealtimePCR与Westernblot检测表明,干旱胁迫导致Rs AQP1b基因与蛋白的表达水平整体下降。PEG6000干旱处理下的转基因与野生型烟草MDA含量都增加,但是转基因烟草的增加幅度小;随着干旱程度加重,转Rs AQP1b基因烟草中SOD含量呈上升趋势,CAT含量呈先上升后下降趋势,POD含量呈上升趋势,酶活性均高于相应野生型对照。
[Abstract]:The full length of RS AQP1b coding sequence of radish aquaporin gene was obtained by homologous cloning from the radish variety Bai Yuchun. The expression pattern of RS AQP1b gene and protein was analyzed under drought stress at seedling stage of radish, and the gene was expressed in tobacco. The changes of MDA content and protective enzyme activity in tobacco under drought stress were studied. The results showed that the encoding sequence of radish aquaporin gene RS AQP1b was 861 BP, encoding 286 amino acids. The polyclonal antibody to RS AQP1b was prepared. Realtime PCR and Westernblot analysis showed that the expression level of Rs AQP1b gene and protein decreased under drought stress. The MDA content of transgenic and wild-type tobacco increased under drought treatment, but the increase of transgenic tobacco was small. With the increase of drought degree, the content of SOD in transgenic AQP1b transgenic tobacco showed an increasing trend, and the content of cat increased first and then decreased, and the activity of enzyme was higher than that of the corresponding wild-type control.
【作者单位】：廊坊师范学院生命科学学院;
【基金】：河北省教育厅青年基金项目(QN2017351) 廊坊师范学院校级项目(LSLQ201407) 廊坊师范学院教改项目(K2015-13)
【分类号】：S631.1

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