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烟草Sar8.2b基因启动子的克隆与鉴定,该基因参与系统获得性抗性。

Cloning and identification of the promoter of the tobacco Sar8.2b gene, a gene involved in systemic acquired resistance.

作者信息

Song Fengming, Goodman Robert M

机构信息

Department of Plant Pathology, University of Wisconsin, Russell Laboratories, Room 689, 1630 Linden Drive, Madison, WI 53706, USA.

出版信息

Gene. 2002 May 15;290(1-2):115-24. doi: 10.1016/s0378-1119(02)00584-x.

Abstract

Expression of the Sar8.2 gene family is induced by salicylic acid (SA) in tobacco during induction of systemic acquired resistance. Expression of Sar8.2b, one member of this 12-member family, was detected as early as 12 h after treatment with SA and was maximal 36 h after SA treatment. In NahG transgenic tobacco plants, benzothiadiazole and dichloroisonicotinic acid induced expression of Sar8.2b but SA did not, suggesting that expression of the Sar8.2b gene is SA-dependent. Several putative cis-acting elements were found in the Sar8.2b gene promoter region, including an as-1 element and GT-1 and Dof binding sequences. We constructed a series of progressive deletion mutations in the Sar8.2b promoter region linked to the beta-glucuronidase (GUS) coding region and analyzed GUS activities by stable expression in transformants of Arabidopsis thaliana. Deletions between -728 and -927 bp or between -351 and -197 bp of the promoter region resulted in a significant reduction in GUS activity induced by SA treatment as shown in stable transformants of A. thaliana. The -197 bp fragment of the promoter region was found to confer a relatively low level of GUS activity induced by SA treatment in stable expression of transformants in A. thaliana. The results suggest that 927 bp of the Sar8.2b gene promoter confers full promoter activity and that cis-acting elements required for high-level SA-inducible expression of the Sar8.2b gene may exist within the regions -728 to -927 bp and -197 to -351 bp.

摘要

在烟草诱导系统获得性抗性过程中,水杨酸(SA)可诱导Sar8.2基因家族的表达。该家族有12个成员,其中Sar8.2b的表达最早在SA处理后12小时即可检测到,在SA处理后36小时达到最大值。在NahG转基因烟草植株中,苯并噻二唑和二氯异烟酸可诱导Sar8.2b的表达,但SA不能,这表明Sar8.2b基因的表达依赖于SA。在Sar8.2b基因启动子区域发现了几个推定的顺式作用元件,包括一个as-1元件以及GT-1和Dof结合序列。我们构建了一系列与β-葡萄糖醛酸酶(GUS)编码区相连的Sar8.2b启动子区域的渐进缺失突变体,并通过在拟南芥转化体中的稳定表达分析了GUS活性。如拟南芥稳定转化体所示,启动子区域-728至-927 bp之间或-351至-197 bp之间的缺失导致SA处理诱导的GUS活性显著降低。在拟南芥转化体的稳定表达中,发现启动子区域的-197 bp片段赋予SA处理诱导的相对较低水平的GUS活性。结果表明,Sar8.2b基因启动子的927 bp赋予了完整的启动子活性,并且Sar8.2b基因高水平SA诱导表达所需的顺式作用元件可能存在于-728至-927 bp和-197至-351 bp区域内。

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