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METHOD FOR DETECTION OF THE ACTIVITY OF GAMMA-SECRETASA.
METHOD FOR DETECTION OF THE ACTIVITY OF GAMMA-SECRETASA.
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机译：检测伽玛-秘法活性的方法。
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摘要
A method for the detection of gamma-secretase activity, in which A. a transgene encoding a fusion protein is used comprising: a) a first nucleotide sequence encoding a protein containing the amino acid sequence GAIIGLMVGGVVIATVIVITLVML ( SEQ ID No. 1), wherein said protein can be cleaved within said amino acid sequence by gamma-secretase present in a cell, thereby forming a first partial protein, which contains the amino acid sequence GAIIGLMVGGVV (SEQ ID No. 2), and a second partial protein, which contains the amino acid sequence VIVI TLVML (SEQ ID No. 3), b) at the 5 '' end of the first nucleotide sequence, a second nucleotide sequence encoding a signal peptide , c) a promoter and d) additional coding nucleotide sequences encoding a protein that is expressed as a fusion protein with the first partial protein and according to it gives partial protein and is used for the detection of the second partial protein and, if appropriate, additional non-coding nucelotide sequences; B. This transgene is incorporated into a human cell or a non-human eukaryotic cell and the fusion protein is expressed; C. the fusion protein is cleaved within the amino acid sequence SEQ ID No. 1 by gamma-secretase present in the cell, whereby the first partial protein and the second partial protein are formed, and D. the second protein is detected partial, characterized in that with the exception of SEQ ID No. 1 that said fusion protein does not contain one or more peptide motifs that act as a signal for endocytosis or exocytosis and / or the protease cleavage site.
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机译：一种用于检测γ-分泌酶活性的方法，其中，使用编码融合蛋白的转基因，其包括：a）编码包含氨基酸序列GAIIGLMVGGVVIATVIVITLVML（SEQ ID No.1）的蛋白质的第一核苷酸序列，其中蛋白可以通过细胞中存在的γ-分泌酶在所述氨基酸序列内切割，从而形成包含氨基酸序列GAIIGLMVGGVV（SEQ ID No.2）的第一部分蛋白和包含氨基酸的第二部分蛋白。酸序列VIVI TLVML（SEQ ID No.3），b）位于第一个核苷酸序列5''末端的第二个核苷酸序列，编码信号肽的第二个核苷酸序列，c）启动子和d）编码以下蛋白的其他编码核苷酸序列：以与第一部分蛋白的融合蛋白的形式表达，并据此得到部分蛋白，并用于检测第二部分蛋白，如果合适，还用于检测其他非编码核苷酸序列; B.将该转基因掺入人细胞或非人真核细胞中并表达融合蛋白。 C.融合蛋白被存在于细胞中的γ-分泌酶切割在氨基酸序列SEQ ID No.1中，由此形成第一部分蛋白和第二部分蛋白，并且D.第二种蛋白被部分检测到，表征除了SEQ ID No.1之外，所述融合蛋白不包含一个或多个肽基序，所述肽基序充当用于内吞或胞吐和/或蛋白酶切割位点的信号。
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