运用噬菌体随机十二肽库筛选与内毒素结合的高亲 - 第三军医大学学报.doc

文章编号：200804166 运用噬菌体肽库摘要噬菌体肽库中筛选与结合的肽序列并进行鉴定以lipopolysaccharide，LPS）为靶分子对噬菌体随机十二肽库进行4轮亲和筛选获得与LPS结合的噬菌体克隆应用结合实验和克隆斑抑制实验进一步确证。挑选结合力强的克隆进行DNA测序，推导出呈现的多肽序列应用软件进行多肽序列分析。4轮亲和筛选从噬菌体随机肽库中获得了个克隆，挑选1个结合力强的克隆进行DNA序列测序及生物信息学分析噬菌体推导出呈现的多肽序列为HWQWPHWSPPP命名为P11肽检索数据库序列。通过对噬菌体随机肽库的淘选与结合的多肽为进一步提供了实验依据和结构基础。关键词内毒素；噬菌体随机肽库；多肽；生物信息学peptides binding to LPS screened from phage display random peptide library YANG Hai-jie, DUAN Guang-jie, LIU You-sheng*, CHEN Rui, YU De-wen，YANG YuInstitute of pathology, Southwest Hospital, the Third Military Medical University, Chongqing 400038, P.R.China) Abstract: Objective To screen and identify peptides with high affinity to lipopolysaccharide (LPS) from phage display random peptide library. Methods Using LPS as a ligand, phage clones that could bind to LPS were screened from phage display random 12-mer peptide library through 4 rounds of biopanning. After measurement of binding activity by ELISA and plaque reduction test, the phage clones presented high affinity to LPS was selected and amplified. Their DNA were extracted for sequencing, and the coding peptide sequences were deduced from the results and analyzed by bioinformatics including homology analysis through Blast. Results After 4 rounds of effective screen, 86 eluted clones were examined and amino acid sequence of 12 positive clones which present high affinity to LPS were selected and identified. The core peptide sequence of phage clones was deduced as HWQWPHWSPPP (Named P11) through comparing with the results screened from phage display random 7-mer peptide library, which was considered to contain the motif of the common amino acid sequence for LPS affinity. There is no patent declared based on P11 sequences retrieving from patent database and 908 proteins matching partly the peptide were found by homology retrieval. Conclusion Peptides with high affinity to LPS was obtained from phage display random peptide library, which provide us some new laboratory evidence and structural foundation