国家医药技术创新博士基金(96-901-11-033)及武汉市晨光计划(965001037-24)资助项目.
对3′-RACE方法进行了修改,采用一条特异性引物和一条通用Oligo dT引物成功地克隆了东亚钳蝎BmKIT3 3′cDNA末端,与常规3′-RACE方法相比,两步PCR方法具有节约成本、节省时间、增加反应特异性等优点,尤其适用于生物活性肽基因的快速克隆和鉴定.
A modified method of 3′-RACE (rapid amplification of 5′- cDNA ends) can rapidly amplify 3′-end of a cDNA only by using a specific primer and Oligo dT.Contrast with the typical 3′-RACE,two-step PCR not only have the advantages of less cost and shorter experimental time ,but can also improve specificity of PCR. This method especially fits for rapid cloning and characterization of the genes encoding the peptides with biological activity.
朱顺义,李文鑫.两步PCR快速扩增东亚钳蝎BmKIT3 3′cDNA末端序列[J].生物化学与生物物理进展,2000,27(1):94-96
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