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海带配子体光捕获蛋白基因lhcf6的启动子克隆与鉴定

DOI: 10.7668/hbnxb.2014.01.012, PP. 62-68

Keywords: 海带,配子体,lhcf6,启动子,GFP

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Abstract:

根据海带配子体光捕获蛋白基因lhcf6的cDNA全长序列(GenBank登录号:DQ250739)设计引物,利用基因组步移技术克隆得到其在雌、雄配子体中的5’-侧翼序列,长度分别为932,901bp。测序结果比对显示二者相似度为33.3%,说明该基因在雌、雄配子体中具有不同的上游调控区。网络服务器PlantCARE和Softberry启动子元件预测结果表明,二者均含有TATA框、节律性相关元件MBS、光反应相关元件MNF1和GAG-motif,其中雌配子体lhcf6基因5’-侧翼序列还特有LHC基因特征元件AT-rich序列,雄配子体特有节律性相关元件Circadian元件,这些元件可能与该基因表达受光质、光强、节律性等因素的调控有关。将其与GFP报告基因一起构建融合表达载体pCAMBIA1304-Flhcf6和pCAMBIA1304-Mlhcf6,利用pCAMBIA1304作对照,通过电击转化法分别转化莱茵衣藻。激光共聚焦荧光显微镜检测结果显示,含pCAMBIA1304-Flhcf6和pCAMBIA1304-Mlhcf6的转基因藻体中GFP基因均有表达,说明克隆得到的海带雌、雄配子体lhcf6基因的5’-侧翼序列具有启动子功能。

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