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首页> 美国卫生研究院文献>World Journal of Gastroenterology >Screening for PreS specific binding ligands with a phage displayed peptides library
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Screening for PreS specific binding ligands with a phage displayed peptides library

机译：用噬菌体展示的肽库筛选PreS特异性结合配体

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AIM: To construct a random peptide phage display library and search for peptides that specifically bind to the PreS region of hepatitis B virus (HBV).METHODS: A phage display vector, pFuse8, based on the gene 8 product (pVIII) of M13 phage was made and used to construct a random peptide library. E.coli derived thioredoxin-PreS was purified with Thio-bond beads, and exploited as the bait protein for library screening. Five rounds of bio-panning were performed. The PreS-binding specificities of enriched phages were characterized with phage ELISA assay.RESULTS: A phage display vector was successfully constructed as demonstrated to present a pVIII fused HBV PreS1 epitope on the phage surface with a high efficiency. A cysteine confined random peptide library was constructed containing independent clones exceeding 5±108 clone forming unit (CFU). A pool of phages showing a PreS-binding specificity was obtained after the screening against thio-PreS with an enrichment of approximately 400 times. Five phages with high PreS-binding specificities were selected and characterized. Sequences of the peptides displayed on these phages were determined.CONCLUSION: A phage library has been constructed, with random peptides displaying as pVIII-fusion proteins. Specific PreS-binding peptides have been obtained, which may be useful for developing antivirals against HBV infection.
机译：目的：构建随机的肽噬菌体展示文库，并寻找与乙型肝炎病毒（HBV）的PreS区特异性结合的肽。方法：基于M13噬菌体的基因8产物（pVIII）的噬菌体展示载体pFuse8。制备并用于构建随机肽文库。用硫键珠纯化大肠杆菌衍生的硫氧还蛋白-PreS，并用作诱饵蛋白进行文库筛选。进行了五轮生物淘选。结果：成功构建了噬菌体展示载体，证明在噬菌体表面高效表达pVIII融合的HBV PreS1抗原决定簇。构建了一个半胱氨酸受限的随机肽库，该库包含超过5±10 8 个克隆形成单位（CFU）的独立克隆。在以大约400倍的富集度对硫代-PreS进行筛选后，获得了显示PreS结合特异性的噬菌体库。选择并表征了五种具有高PreS结合特异性的噬菌体。结论：构建了噬菌体文库，其中随机肽为pVIII融合蛋白。已获得特定的PreS结合肽，可用于开发抗HBV感染的抗病毒药。

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