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The complete sequence of soybean chlorotic mottle virus DNA and the identification of a novel promoter.

机译:大豆褪绿斑驳病毒DNA的完整序列和新启动子的鉴定。

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摘要

The complete nucleotide sequence of an infectious clone of soybean chlorotic mottle virus (SoyCMV) DNA was determined and compared with those of three other caulimoviruses, cauliflower mosaic virus (CaMV), carnation etched ring virus and figwort mosaic virus. The double-stranded DNA genome of SoyCMV (8,175 bp) contained nine open reading frames (ORFs) and one large intergenic region. The primer binding sites, gene organization and size of ORFs were similar to those of the other caulimoviruses, except for ORF I, which was split into ORF Ia and Ib. The amino acid sequences deduced from each ORF showed only short, highly homologous regions in several of the corresponding ORFs of the three other caulimoviruses. A promoter fragment of 378 bp in SoyCMV ORF III showed a strong expression activity, comparable to that of the CaMV 35S promoter, in tobacco mesophyll protoplasts as determined by a beta-glucuronidase assay using electrotransfection. The fragment contained CAAT and TATA boxes but no transcriptional enhancer signal as reported for the CaMV 35S promoter. Instead, it had sequences homologous to a part of the translational enhancer signal reported for the 5'-leader sequence of tobacco mosaic virus RNA.
机译:确定了大豆绿叶斑驳病毒(SoyCMV)DNA传染性克隆的完整核苷酸序列,并将其与其他三种花椰菜花叶病毒,花椰菜花叶病毒(CaMV),康乃馨蚀刻环病毒和非花草花叶病毒的核苷酸序列进行了比较。 SoyCMV的双链DNA基因组(8,175 bp)包含9个开放阅读框(ORF)和1个较大的基因间区。 ORF的引物结合位点,基因组织和大小与其他花椰菜花叶病毒相似,但ORF I分为ORF Ia和Ib。从每个ORF推导的氨基酸序列在其他三种花椰菜花叶病毒的几个相应的ORF中仅显示出短的,高度同源的区域。 SoyCMV ORF III中378 bp的启动子片段在烟草叶肉原生质体中显示出与CaMV 35S启动子相当的强表达活性,这是通过使用电转染的β-葡萄糖醛酸苷酶测定法确定的。该片段包含CAAT和TATA框,但没有CaMV 35S启动子报道的转录增强子信号。相反,它具有与烟草花叶病毒RNA 5'-前导序列报道的翻译增强子信号一部分同源的序列。

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