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玄参花叶病毒强启动子的特性:与花椰菜花叶病毒类似的35S启动子及调控甘露碱合成酶启动子的比较。

Characteristics of a strong promoter from figwort mosaic virus: comparison with the analogous 35S promoter from cauliflower mosaic virus and the regulated mannopine synthase promoter.

作者信息

Sanger M, Daubert S, Goodman R M

机构信息

Calgene Inc., Davis, CA 95616.

出版信息

Plant Mol Biol. 1990 Mar;14(3):433-43. doi: 10.1007/BF00028779.

Abstract

A segment of DNA from the genome of figwort mosaic virus (FMV) strain M3 possesses promoter activity when tested in electroporated protoplasts from, and transgenic plants of, Nicotiana tabacum cv. Xanthi nc. The 1.1 kb DNA segment, designated the '34S' promoter, is derived from a position on the FMV genome comparable to the position on the cauliflower mosaic virus (CaMV) genome containing the 35S promoter. The 34S and 35S promoters show approximately 63% nucleotide homology in the TATA, CCACT, and -18 to +1 domains, but in sequences further upstream the homology drops below 50%. Promoter activities were estimated using beta-glucuronidase and neomycin phosphotransferase II reporter gene systems. The activity of the 34S promoter segment approximates that of the 35S promoter in both protoplast transient expression assays and in stably transformed tobacco plants. Truncation of 5' sequences from the 34S promoter indicates that promoter strength depends upon DNA sequences located several hundred nucleotides upstream from the TATA box. In leaf tissue the 34S promoter is 20-fold more active than the mannopine synthase (MAS) promoter from Agrobacterium tumefaciens T-DNA. The 34S promoter lacks the root-specific and wound-stimulated expression of the MAS promoter, showing relatively uniform root, stem, leaf, and floral activities.

摘要

在对烟草品种Xanthi nc的电穿孔原生质体和转基因植株进行测试时,玄参花叶病毒(FMV)M3株系基因组中的一段DNA具有启动子活性。这个1.1 kb的DNA片段被命名为“34S”启动子,它来源于FMV基因组上一个与花椰菜花叶病毒(CaMV)基因组中含有35S启动子的位置相当的位点。34S和35S启动子在TATA、CCACT以及-18至+1区域显示出约63%的核苷酸同源性,但在更上游的序列中,同源性降至50%以下。使用β-葡萄糖醛酸酶和新霉素磷酸转移酶II报告基因系统评估启动子活性。在原生质体瞬时表达试验和稳定转化的烟草植株中,34S启动子片段的活性与35S启动子相近。从34S启动子截断5'端序列表明,启动子强度取决于位于TATA框上游数百个核苷酸处的DNA序列。在叶片组织中,34S启动子的活性比根癌农杆菌T-DNA的甘露碱合成酶(MAS)启动子高20倍。34S启动子缺乏MAS启动子的根特异性和伤口刺激诱导表达,在根、茎、叶和花中表现出相对均匀的活性。

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