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目的:建立人类K-ras基因突变质控品,用于K-ras基因突变检测试剂盒的注册检测工作。方法:根据人类K-ras基因序列设计引物,通过定点突变技术引入突变位点,将获得的PCR产物与克隆载体连接,转化感受态细菌后筛选阳性克隆,获得了包含不同突变位点的重组质粒,建立了人类K-ras基因突变质控品。结果:对人类K-ras基因突变质控品进行序列测定,并将测序结果进行BLAST比对,结果表明成功建立了人类K-ras基因突变质控品。结论:建立的K-ras基因突变体质控品包括常见的Kras基因12密码子突变类型,可以用于指导Kras基因突变检测试剂盒的注册检验工作。
Objective: To establish a human K-ras gene mutation control product for K-ras gene mutation detection kit registration testing. Methods: The primers were designed according to the human K-ras gene sequence. Mutations were introduced by site-directed mutagenesis. The PCR products were ligated with the cloning vector and transformed into competent bacteria. The positive clones were selected and the recombinant plasmids containing different mutation sites were obtained , Established a human K-ras gene mutation quality control. Results: The sequence of human K-ras gene mutation was sequenced and compared with BLAST. The results showed that human K-ras gene mutation control was successfully established. CONCLUSION: The K-ras mutants contain the common Kras mutant codon 12 codon and can be used to guide the registration of Kras gene mutation detection kit.