噬菌体展示随机肽克隆的一种快速、高通量测序技术  

A rapid and high throughput sequencing method of the phage display random peptide in a murine model of asthma

作　　者：杨芳[1] 张文红[2] 

机构地区：[1]安康学院农学与生命科学院,陕西安康725000 [2]第四军医大学基因诊断技术应用研究所,陕西西安710032

出　　处：《东南大学学报（医学版）》2011年第3期453-455,共3页Journal of Southeast University(Medical Science Edition)

基　　金：陕西省教育厅自然科学基金资助项目(08JK210);安康学院高层次人才科研专项(AYQDZR200805)

摘　　要：目的:探索应用焦磷酸微测序(Pyrosequencing)技术建立一种针对噬菌体展示随机肽的高通量序列测定方案。方法:环7肽噬菌体展示随机肽库的淘筛后随机挑选10个蓝色噬菌斑作为模板,PCR扩增随机7肽区序列,应用Pyrosequencing技术进行实时测序和分析。结果:应用Pyrosequencing技术对10例样品的随机7肽区进行序列分析,得到一个GXXXHPQ的同源基序(X为任意氨基酸),此基序为链亲合素的结合肽基序,结果与预期相符合。结论:Pyrosequencing技术具有操作简易快速、高通量的优点,必定可以广泛应用于各种随机肽库的筛选测序。Objective： To develop a pyrosequencing-based method for selection of the phage display random peptide.Methods： After selection of Ph.D.-C7C phage display peptide library and PCR amplification of random gene sequence of Ph.D.-C7C,the PCR products were further sequenced by pyrosequencing.Results： The sequence of Ph.D.-C7C PCR products was identified by pyrosequencing,the regions were highly homologous with the motifs of GXXXHPQ,which coincided with the sequence of peptide binding to streptavidin.Conclusion： Pyrosequencing used in the analysis of the phage display random peptide has the advantage of high resolution and can be widely employed in sequencing of the phage display random peptide library.

关 键 词：随机肽 测序 焦磷酸微测序技术 

分 类 号：Q503[生物学—生物化学] Q782