Keil M., Sanchez-Serrano J., Schell J., Willmitzer L.
Institut fur Genbiologische Forschung Berlin GmbH, Ihnestrasse 63, D-1000 Berlin 33, Federal Republic of Germany.
Plant Cell. 1990 Jan;2(1):61-70. doi: 10.1105/tpc.2.1.61.
The effect of progressive 5[prime] deletions within a potato proteinase inhibitor II promoter on wound-inducible expression of the chloramphenicol acetyltransferase (CAT) gene in leaves of transgenic tobacco plants was analyzed. After deletion of a region ranging from position -1300 to -700 with respect to the transcription start site, promoter activity was markedly reduced but still wound-inducible. Further deletion of approximately 200 base pairs resulted in a promoter activity that was below the detection limit, proving that the activity of the proteinase inhibitor II promoter is controlled by sequences upstream of position -514. Addition of the enhancer of the 35S promoter of the cauliflower mosaic virus (CaMV) either 5[prime] upstream or 3[prime] downstream of chimeric genes consisting of different proteinase inhibitor II promoter deletions (-700, -514, -210) fused to the CAT gene led to wound-inducible CAT gene expression in a fraction of transgenic plants containing either the "-700" or "-514" promoters, indicating the presence of wound-responsive elements in the promoter-proximal region. A fragment of the proteinase inhibitor II promoter comprising sequences between positions -1300 and -195 is able to confer wound-inducible expression to an inactive CaMV 35S promoter truncated at position -90 in either orientation, proving that this fragment displays wound-specific, enhancer-like properties. In addition, data are presented excluding that the proteinase inhibitor II 3[prime] end is of importance for wound-inducible gene expression.
分析了马铃薯蛋白酶抑制剂II启动子内5′端逐步缺失对转基因烟草植株叶片中氯霉素乙酰转移酶(CAT)基因伤口诱导表达的影响。相对于转录起始位点缺失从-1300至-700位置的区域后,启动子活性显著降低,但仍具有伤口诱导性。进一步缺失约200个碱基对导致启动子活性低于检测限,证明蛋白酶抑制剂II启动子的活性受-514位置上游序列的控制。在由不同蛋白酶抑制剂II启动子缺失(-700、-514、-210)与CAT基因融合而成的嵌合基因的5′上游或3′下游添加花椰菜花叶病毒(CaMV)35S启动子的增强子,导致在含有“-700”或“-514”启动子的一部分转基因植株中出现伤口诱导的CAT基因表达,表明在启动子近端区域存在伤口响应元件。包含-1300至-195位置之间序列的蛋白酶抑制剂II启动子片段能够赋予在-90位置截短的无活性CaMV 35S启动子以伤口诱导表达,无论其方向如何,证明该片段具有伤口特异性的、类似增强子的特性。此外,所提供的数据排除了蛋白酶抑制剂II 3′端对伤口诱导基因表达的重要性。