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Split内含肽高效介导转录激活因子的胞内融合 被引量：1

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摘要 Split内含肽可以通过反式剪接作用介导两个蛋白片段的共价连接.基于这一原理,利用来自于Synechocystis sp.strain PCC6803的天然split内含肽DnaEs作为蛋白剪接元件,通过其反式剪接作用介导了四环素阻遏蛋白(TetR)与1-型单纯疱疹病毒(HSV)的转录活化域(VP16)的共价连接,形成融合蛋白TV.应用RT-PCR技术以及荧光显微镜成像技术分别在mRNA转录水平与蛋白表达水平上研究了DnaEs的剪接活性,结果证明在细胞内DnaEs可以介导融合蛋白TV的产生.同时也对融合蛋白TV的转录活性进行了研究,并将其与通过重组DNA表达得到的同种蛋白的转录活性进行了比较,发现DnaEs介导的融合蛋白与基因重组得到的同种蛋白转录活性相似.此外,还研究了四环素类似物Dox对TV转录活性的调控,结果表明Dox可以明显抑制其转录活性.DnaEs介导转录激活因子融合,并启动下游报告基因表达的实验方法,为深入开展DnaEs的应用研究提供了重要实验依据.
作者 高元吉 赵传科 尹琪 王慧 曾新 王少鹏 于文文 栗世铀
机构地区 中国科学院北京基因组研究所新药研发组 中国科学院植物研究所
出处 《中国科学（C辑）》 CSCD 北大核心 2008年第8期714-721,共8页 Science in China(Series C)
关键词 DnaEs 蛋白剪接 四环素阻遏蛋白 1-型单纯疱疹病毒 转录活化域 转录激活因子
分类号 Q75 [生物学—分子生物学]
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参考文献30

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同被引文献22

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中国科学（C辑）
2008年 第8期

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