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植物G盒启动子序列可在酿酒酵母中激活转录,并且在体外可被一种类似于植物G盒结合因子GBF的酵母活性物质所结合。

The plant G box promoter sequence activates transcription in Saccharomyces cerevisiae and is bound in vitro by a yeast activity similar to GBF, the plant G box binding factor.

作者信息

Donald R G, Schindler U, Batschauer A, Cashmore A R

机构信息

Department of Biology, University of Pennsylvania, Philadelphia 19104.

出版信息

EMBO J. 1990 Jun;9(6):1727-35. doi: 10.1002/j.1460-2075.1990.tb08296.x.

Abstract

G box and I box sequences of the Arabidopsis thaliana ribulose-bisphosphate-1,5-carboxylase small subunit (RBCS) promoter are required for expression mediated by the Arabidopsis rbcS-1A promoter in transgenic tobacco plants and are bound in vitro by factors from plant nuclear extracts termed GBF and GA-1, respectively. We show here that a -390 to -60 rbcS-1A promoter fragment containing the G box and two I boxes activates transcription from a truncated iso-1-cytochrome c (CYC1) gene promoter in Saccharomyces cerevisiae. Mutagenesis of either the rbcS-1A G box or both I box sequences eliminated the expression mediated by this fragment. When polymerized, I box oligonucleotides were also capable of enhancing expression from the truncated CYC1 promoter. Single-copy G box sequences from the Arabidopsis rbcS-1A, Arabidopsis Adh and tomato rbcS-3A promoters were more potent activators and were used in mobility shift assays to identify a DNA binding activity in yeast functionally similar to GBF. In methylation interference experiments, the binding specificity of the yeast protein was indistinguishable from that obtained with plant nuclear extracts.

摘要

拟南芥1,5 - 二磷酸核酮糖羧化酶小亚基(RBCS)启动子的G盒和I盒序列,对于转基因烟草植株中由拟南芥rbcS - 1A启动子介导的表达是必需的,并且在体外分别与来自植物核提取物的称为GBF和GA - 1的因子结合。我们在此表明,一个包含G盒和两个I盒的-390至-60 rbcS - 1A启动子片段,可激活酿酒酵母中截短的异-1-细胞色素c(CYC1)基因启动子的转录。对rbcS - 1A G盒或两个I盒序列进行诱变,可消除该片段介导的表达。当I盒寡核苷酸聚合时,也能够增强截短的CYC1启动子的表达。来自拟南芥rbcS - 1A、拟南芥Adh和番茄rbcS - 3A启动子的单拷贝G盒序列是更强的激活剂,并用于迁移率变动分析,以鉴定酵母中与GBF功能相似的DNA结合活性。在甲基化干扰实验中,酵母蛋白的结合特异性与植物核提取物的无异。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e0d/551875/902c9bc66c18/emboj00233-0057-a.jpg

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