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Cap-independent translation sequences derived from barley yellow dwarf virus

机译:大麦黄矮病毒衍生的不依赖帽的翻译序列

摘要

The present invention provides a method of increasing the production of a protein translated from an uncapped eukaryotic messenger ribonucleic acid (mRNA), comprising the steps of: selecting a nucleotide sequence encoding a protein to be expressed; joining nucleotides 1-168 of the barley yellow dwarf virus RNA, PAV serotype to the 5' untranslated region of said uncapped mRNA, said nucleotides 1-168 comprising the 5' untranslated region plus first 27 nucleotides of the open reading frame of the barley yellow dwarf virus RNA; linking nucleotides 4513-5677 of the barley yellow dwarf virus RNA, PAV serotype, to the 3' untranslated region of said uncapped mRNA; and expressing the protein. Also provided is a DNA molecule which comprises: (a) a promoter region which functions in plant cells to cause the production of an RNA sequence, which is operably linked to (b) a 5' untranslated region including a 5' translation enhancing segment, said 5' translation enhancing segment comprising nucleotides 1-168 of the barley yellow dwarf virus RNA, PAV serotype, said nucleotides 1-168 encompassing the 5' untranslated region plus first 27 nucleotides of the open reading frame of the barley yellow dwarf virus RNA; (c) a coding sequence, wherein said coding sequence is heterologous to said 5' untranslated region, which is operably linked to (d) a 3' untranslated region that functions in plant cells to cause the termination of transcription, said 3' untranslated region comprising nucleotides 4513-5677 of the barley yellow dwarf virus RNA.
机译:本发明提供了一种增加从无盖真核信使核糖核酸(mRNA)翻译的蛋白质的产量的方法,该方法包括以下步骤:选择编码待表达蛋白质的核苷酸序列;和将大麦黄矮病毒RNA,PAV血清型的核苷酸1-168与所述未封端的mRNA的5'非翻译区连接,所述核苷酸1-168包括5'非翻译区加上大麦黄开放阅读框的前27个核苷酸矮病毒RNA;将大麦黄矮病毒RNA PAV血清型的核苷酸4513-5677连接至所述未封端的mRNA的3'非翻译区;并表达蛋白质。还提供了一种DNA分子,其包含:(a)在植物细胞中起作用以引起RNA序列产生的启动子区,该启动子区与(b)包括5'翻译增强区段的5'非翻译区可操作地连接,所述5'翻译增强片段,包含大麦黄矮病毒RNA,PAV血清型的核苷酸1-168,所述核苷酸1-168涵盖了大麦黄矮病毒RNA的5'非翻译区和开放阅读框的前27个核苷酸; (c)编码序列,其中所述编码序列与所述5'非翻译区是异源的,其可操作地连接至(d)在植物细胞中起作用以引起转录终止的3'非翻译区,所述3'非翻译区包含大麦黄矮病毒RNA的核苷酸4513-5677。

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