公开(公告)号：US5910628A

公开(公告)日：1999-06-08

申请号：US858623

申请日：1997-05-20

申请人： W. Allen Miller ,  Shanping Wang

发明人： W. Allen Miller ,  Shanping Wang

IPC分类号： C12N15/67 ,  C12N15/81 ,  C12N15/82 ,  C12N15/83 ,  A01H1/04

CPC分类号： C12N15/67 ,  C12N15/81 ,  C12N15/8216

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.

摘要翻译： 本发明提供了一种增加从未加帽的真核信使核糖核酸（mRNA）翻译的蛋白质的产生的方法，包括以下步骤：选择编码待表达蛋白质的核苷酸序列; 将大麦黄矮病毒RNA，PAV血清型的核苷酸1-168连接到所述未加帽的mRNA的5'非翻译区，所述核苷酸1-168包含大麦黄开放阅读框的5'非翻译区加上第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连接到所述未加帽的mRNA的3'非翻译区; 并表达蛋白质。 还提供了DNA分子，其包含：（a）在植物细胞中起作用以产生RNA序列的启动子区，其可操作地连接到（b）包含5'翻译增强区的5'非翻译区， 所述5'翻译增强区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包含大麦黄矮病毒RNA的开放阅读框的5'非翻译区加上第27个核苷酸; （c）编码序列，其中所述编码序列与所述5'非翻译区异源，其可操作地连接到（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 包括大麦黄矮病毒RNA的核苷酸4513-5677。