随机序列八肽库的构建及其在双杂交系统中的应用

Construction of Random Eight-peptide Library and Its Use in Yeast Two-hybrid System

目的:构建一个随机序列八肽库并从中寻找非天然的具有药用价值的多肽。方法:利用基因工程技术构建随机序列八肽库,首先设计并合成了含8个密码子的随机寡核苷酸链,摸索了变性温度与时间、复性温度与时间、延伸温度与时间以及循环数后利用PCR扩增了这一片段,并解决了片段回收、连接等步骤,最后比较了不同的转化方法,采用最简便高效的TSS法转化大肠杆菌建立了随机序列八肽库。结果:利用作者所建立的随机序列八肽库应用于酵母双杂交系统并从中筛选到了一段感兴趣的多肽。结论:随机序列八肽库构建成功并且具有应用价值。

Objective：Construction of a random peptide library to find non-natural produced polypeptide which can be a new drug candidate.Methods： The random eight-peptide library was constructed by genetic engineering.First,a random oligonucleotides fragment containing eight codons was designed and synthesized.The fragment was amplified by PCR after the optimal denaturing temperature and time,annealing temperature and time,elongation temperature and time,cycles were determined.Then,the recovery and ligation of this small fragment were also studied.Finally, different trnsformation methods were compared, among which TSS, the most efficient method, was selocted to construct the random eight - peptide library in E. coli. Results： A poem fragment was screened out from the cstablished random eight- peptide library by using yeast two - hybrid syserm. Conclusion： The random eight - peptide was successfully and established and workable.