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METHOD FOR DETECTING NONSENSE MUTATIONS AND FRAMESHIFT MUTATIONS
METHOD FOR DETECTING NONSENSE MUTATIONS AND FRAMESHIFT MUTATIONS
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机译：检测无意义变异和框架变异的方法
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摘要
A method for detecting nonsense mutations and frameshift mutations which is simple and which may be applied even when the size of the test DNA is large is disclosed. In the method of the present invention, a test nucleic acid fragment is inserted into a site of a vector having a promoter, a translational initiation codon downstream of the promoter, a reporter gene which is a structural gene located downstream of the translational initiation codon, which is operably linked to the promoter and which encodes a polypeptide. A fusion polypeptide formed by ligating the N-terminal of the polypeptide to another polypeptide is detectable based on a function of the polypeptide encoded by the reporter gene. The site into which the test nucleic acid fragment is inserted is located downstream of the translational initiation codon and upstream of the reporter gene, and the test nucleic acid fragment is one which allows, when inserted, in-frame location of the reporter gene with respect to the translational initiation codon when the test nucleic acid fragment is normal type. The test nucleic acid fragment and the reporter gene downstream thereof in the resulting recombinant vector are then expressed in a host cell, and whether or not the fusion polypeptide having the function of the polypeptide encoded by the reporter gene is produced is determined.
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