Yang I C, Iommarini J P, Becker D K, Hafner G J, Dale J L, Harding R M
Centre for Molecular Biotechnology, Queensland University of Technology, GPO Box 2434, 4001, Brisbane, Australia.
Plant Cell Rep. 2003 Aug;21(12):1199-206. doi: 10.1007/s00299-003-0621-x. Epub 2003 May 15.
Taro bacilliform virus (TaBV) is a pararetrovirus of the genus Badnavirus which infects the monocotyledonous plant, taro ( Colocasia esculenta). A region of the TaBV genome spanning nucleotides 6,281 to 12 (T1200), including the 3' end of open reading frame 3 (ORF 3) and the intergenic region to the end of the tRNA(met)-binding site, was tested for promoter activity along with four different 5' deletion fragments (T600, T500, T250 and T100). In transient assays, only the T1200, T600, T500 fragments were shown to have promoter activity in taro leaf, banana suspension cells and tobacco callus. When these three promoters were evaluated in stably transformed, in vitro-grown transgenic banana and tobacco plants, all were found to drive near-constitutive expression of either the green fluorescent protein or beta-glucuronidase (GUS) reporter gene in the stem (or pseudostem), leaves and roots, with strongest expression observed in the vascular tissue. In transgenic banana leaves, the T600 promoter directed four-fold greater GUS activity than that of the T1200, T500 and the maize polyubiquitin-1 promoters. In transgenic tobacco leaves, the levels of GUS expression directed by the three promoters was between four- and ten-fold lower than that of the double Cauliflower mosaic virus 35S promoter. These results indicate that the TaBV-derived promoters may be useful for the high-level constitutive expression of transgenes in either monocotyledonous or dicotyledonous species.
芋头杆状病毒(TaBV)是杆状DNA病毒属的一种副逆转录病毒,可感染单子叶植物芋头(芋属植物)。对TaBV基因组中跨越核苷酸6281至12(T1200)的区域进行了启动子活性测试,该区域包括开放阅读框3(ORF 3)的3'端以及至tRNA(met)结合位点末端的基因间区域,同时还测试了四个不同的5'缺失片段(T600、T500、T250和T100)。在瞬时分析中,仅T1200、T600、T500片段在芋头叶片、香蕉悬浮细胞和烟草愈伤组织中表现出启动子活性。当在稳定转化的、体外培养的转基因香蕉和烟草植株中评估这三种启动子时,发现它们均能驱动绿色荧光蛋白或β-葡萄糖醛酸酶(GUS)报告基因在茎(或假茎)、叶和根中近乎组成型表达,在维管组织中表达最强。在转基因香蕉叶片中,T600启动子指导的GUS活性比T1200、T500和玉米多聚泛素-1启动子高四倍。在转基因烟草叶片中,这三种启动子指导的GUS表达水平比双花椰菜花叶病毒35S启动子低四至十倍。这些结果表明,源自TaBV的启动子可能有助于在单子叶或双子叶物种中高水平组成型表达转基因。