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METHOD FOR DETECTING NONSENSE MUTATIONS AND FRAMESHIFT MUTATIONS

机译:检测无意义变异和框架变异的方法

摘要

A method for conveniently detecting nonsense mutations and frameshift mutations, which is applicable even to large-sized test DNAs. This method comprises inserting, into a vector containing a promoter, an initiation codon located downstream the promoter and a reporter gene located downstream the initiation codon which is a structural gene under the regulation by the promoter and a fused polypeptide consisting of the polypeptide encoded by which and another polypeptide bonded to the N-terminus of the former polypeptide can be detected based on the function of the polypeptide encoded by the structural gene, a test nucleic acid fragment which makes the reading frame of the reporter gene to match the above-mentioned initiation codon when it is a normal type one to a site downstream the initiation codon but upstream the reporter gene; expressing the test nucleic acid fragment in the recombinant vector thus obtained and the reporter gene downstream the same in host cells; and thus examining whether or not a fused polypeptide having the function of the polypeptide encoded by the reporter gene is produced.
机译:一种方便地检测无义突变和移码突变的方法,该方法甚至适用于大型测试DNA。该方法包括将位于启动子下游的起始密码子和位于起始密码子下游的报告基因插入到包含启动子的载体中,该报告基因是在启动子和由其编码的多肽组成的融合多肽的调控下的结构基因。根据结构基因编码的多肽的功能,可以检测与前一个多肽的N末端键合的另一个多肽,该结构核酸是一种使报告基因的阅读框与上述起始序列相匹配的测试核酸片段。当是起始密码子下游但报告基因上游位点的正常类型的密码子时;在如此获得的重组载体中表达测试核酸片段,并在宿主细胞中表达其下游的报道基因。因此,检查是否产生了具有由报道基因编码的多肽的功能的融合多肽。

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