从噬菌体展示肽库中筛选与人精子蛋白17结合的短肽序列

Screening human sperm protein 17 binding peptides from phage display library

目的:随机从噬菌体12肽库中筛选与人精子蛋白17(Sp17)特异性结合的短肽序列,并进行鉴定。方法:以基因工程制备的人Sp17为钓饵蛋白,采用亲和淘筛法对噬菌体肽库进行三轮亲和淘筛。富集Sp17特异结合噬菌体,用ELISA鉴定其结合活性并进行DNA测序分析。结果:随机挑出20个噬菌体克隆进行鉴定,ELISA显示其中5个克隆与Sp17有较强结合活性,其氨基酸序列分别为QLMSNIHRRM II、RKMMNQTKRHLP、NTMKTMR IMMTR、RRRLLLMQRRMKR和SPRPMMRR IRKQ。结论:从噬菌体展示肽库中筛选鉴定出5个与人Sp17结合的12肽序列。

Objective：To screen and identify peptides that bind specifically to human sperm protein 17 （Sp17） from 12 mer phage display library. Methods：The recombinant Sp17 was exploited as the bait protein for library screening. Three rounds of bio-panning were performed. The Sp17-binding specificities of enriched phages were characterized with ELISA assay. Five phage clones with very high affinity for Sp17 were selected and sequences of the peptides displayed on these phages were determined. Results ： The phages that bind specifically to Sp17 were enriched through 3 rounds of biopanning, 20 clones of phage were randomly picked, of which 5 positive phage clones showed specific binding with Sp17 without binding with unrelated proteins of human serum, deduced amino acid sequences of selected 5 peptides are QLMSNIHRRMII, RKMMNQTKRHLP, NTMKTMRIMMTR, RRRLLLMQRRMKR 和 SPRPMMRRIRKQ. Conclusion：Five different peptides which specifically bind to Sp17 were selected and identified.