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马铃薯创伤诱导型蛋白酶抑制剂II基因3'调控区的功能分析

Functional analysis of the 3' control region of the potato wound-inducible proteinase inhibitor II gene.

作者信息

An G, Mitra A, Choi H K, Costa M A, An K, Thornburg R W, Ryan C A

机构信息

Institute of Biological Chemistry, Washington State University, Pullman 99164-6340.

出版信息

Plant Cell. 1989 Jan;1(1):115-22. doi: 10.1105/tpc.1.1.115.

Abstract

Proteinase inhibitor genes are expressed strongly in specific plant tissues under both developmental and environmental regulation. We have studied the role of the 3' control region of the potato proteinase inhibitor II gene (PI-II) that is inducible in leaves in response to herbivore attacks or other severe wounding. Comparison of the terminator from the PI-II gene with two different terminators from the 6b and 7 genes, driven by a common PI-II promoter-cat fusion molecule, indicated that the PI-II terminator provided the most efficient expression of cat. The PI-II terminator also caused a significantly elevated cat gene expression driven by the cauliflower mosaic virus 35S promoter. The increase in the level of expression is probably not due to the presence of an enhancer element in the PI-II terminator region, but to cis-acting elements involved in mRNA processing or stability. Both transient and stable transformation analyses of the deletion mutants in the 3'-flanking sequence indicated that about a 100-base pair DNA fragment surrounding the polyadenylation site is essential for the efficient gene expression. This region seems to consist of several regulatory elements, including the conserved sequence, CGTGTCTT, which is located 9 bases downstream from the polyadenylation site. The elements appear to contribute to the increased stability of mRNAs containing the PI-II terminator.

摘要

蛋白酶抑制剂基因在发育和环境调控下于特定植物组织中强烈表达。我们研究了马铃薯蛋白酶抑制剂II基因(PI-II)3'调控区的作用,该基因在叶片中可被食草动物攻击或其他严重创伤诱导。由一个共同的PI-II启动子 - cat融合分子驱动,将PI-II基因的终止子与来自6b和7基因的两种不同终止子进行比较,结果表明PI-II终止子使cat表达效率最高。PI-II终止子还导致由花椰菜花叶病毒35S启动子驱动的cat基因表达显著升高。表达水平的增加可能不是由于PI-II终止子区域存在增强子元件,而是由于参与mRNA加工或稳定性的顺式作用元件。对3'侧翼序列缺失突变体的瞬时和稳定转化分析均表明,围绕聚腺苷酸化位点的约100个碱基对的DNA片段对于高效基因表达至关重要。该区域似乎由几个调控元件组成,包括保守序列CGTGTCTT,其位于聚腺苷酸化位点下游9个碱基处。这些元件似乎有助于增加含有PI-II终止子的mRNA的稳定性。

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