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CREB-binding protein transcription activation domain for enhanced transgene expression by a positive feedback system

机译:CREB结合蛋白转录激活域可通过正反馈系统增强转基因表达

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摘要

The positive feedback system using a fusion protein of the sequence-specific DNA binding domain of yeast GAL4 and the transcription activation domain of herpes simplex virus VP16 (GAL4-VP16), in which GAL4-VP16 promotes its own expression as well as that of a reporter gene product, is useful for efficient transgene expression from plasmid DNA. In this study, the transcription activation domains of endogenous proteins, instead of VP16, were fused to the GAL4 DNA binding domain, and the positive feedback systems employing the novel fusion proteins were examined. Plasmid DNAs encoding the transcription factors were introduced into mouse Hepa 1-6 cells by electroporation and lipofection. Among CREB-binding protein (226-460), sterol regulatory element-binding protein-1 (1-140), p53 (1-70), and Med15 (9-73), the CREB-binding protein functioned efficiently as an activator. These results indicated that the GAL4-CREB-binding protein is useful for enhanced transgene expression by the positive feedback system. (C) 2011 Elsevier B.V. All rights reserved.
机译:使用酵母GAL4的序列特异性DNA结合结构域和单纯疱疹病毒VP16(GAL4-VP16)的转录激活结构域的融合蛋白的正反馈系统,其中GAL4-VP16促进其自身表达以及报告基因产物可用于从质粒DNA有效表达转基因。在这项研究中,将内源蛋白的转录激活域而不是VP16与GAL4 DNA结合域融合,并研究了使用新型融合蛋白的正反馈系统。通过电穿孔和脂转染将编码转录因子的质粒DNA引入小鼠Hepa 1-6细胞。在CREB结合蛋白(226-460),固醇调节元件结合蛋白-1(1-140),p53(1-70)和Med15(9-73)中,CREB结合蛋白有效地发挥了激活剂的作用。这些结果表明GAL4-CREB结合蛋白可用于通过正反馈系统增强转基因表达。 (C)2011 Elsevier B.V.保留所有权利。

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