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    • 63. 发明授权
    • Heterologous polypeptide production in the absence of nonsense-mediated
MRNA decay functions
    • 在没有无义介导的MRNA衰变功能的情况下产生异源多肽
    • US6071700A
    • 2000-06-06
    • US177431
    • 1998-10-21
    • Feng HeAllan S. Jacobson
    • Feng HeAllan S. Jacobson
    • C07K14/395C12N1/15C12N15/12C12N15/67C12Q1/68C12N15/31
    • C07K14/395C12N15/67
    • The invention relates to the discovery of a gene, NMD2, named after its role in the Nonsense-Mediated mRNA Decay pathway, and the protein, Nmd2p, encoded by the NMD2 gene. The amino acid sequence of Nmd2p and the nucleotide sequence of the NMD2 gene encoding it are disclosed. Nmd2p is shown herein to bind to another protein in the decay pathway, Upf1p. A C-terminal fragment of the protein is also shown to bind Upf1p and, when overexpressed in the host cell, the fragment inhibits the function of Upf1p, thereby inhibiting the nonsense-mediated mRNA decay pathway. The invention also relates to methods of inhibiting the nonsense-mediated mRNA decay pathway to stabilize mRNA transcripts containing a nonsense codon which normally would cause an increase in the transcript decay rate. Such stabilization of a transcript is useful for the production of a recombinant protein or fragment thereof. The invention also relates to methods of identifying molecules that inhibit the nonsense-mediated mRNA decay pathway, and the use of such molecules for treatment of disorders associated with nonsense mutations.
    • 本发明涉及以其在无义介导的mRNA衰变途径中的作用命名的NMD2基因和由NMD2基因编码的蛋白质Nmd2p的发现。 公开了Nmd2p的氨基酸序列和编码它的NMD2基因的核苷酸序列。 Nmd2p在本文中显示为与衰老途径Upf1p中的另一种蛋白结合。 蛋白质的C末端片段还显示结合Upf1p,并且当在宿主细胞中过表达时,该片段抑制Upf1p的功能,从而抑制无义介导的mRNA衰减途径。 本发明还涉及抑制无义介导的mRNA衰减途径以稳定含有无义密码子的mRNA转录物的方法,其通常将导致转录物衰减率的增加。 转录物的这种稳定化对于重组蛋白质或其片段的产生是有用的。 本发明还涉及鉴定抑制无义介导的mRNA衰变途径的分子的方法,以及这些分子用于治疗与无义突变相关的病症的用途。