应用抗病毒抗体表位筛选的多肽文库的构建

1985年,Sndth在《科学》杂志上提出构建"融合噬菌体",即在这种丝状病毒的包膜蛋白上表达融合蛋白1]．1990年,Scott和Smith山利用这种表面表达载体建立了随机多肽文库,可以很方便地筛选出抗体及其他功能蛋白的强结合配基2]．我们建立了十二肽的随机多肽库,将从中筛选出抗HIV-gP160抗体的抗独特型多肽．1材料和方法1．1质粒、菌株和培养墓噬菌粒成h血四、大肠杆菌XLI-Blue和辅助噬菌体VCSM13由军事医学科学院微生物流行病研究所王海涛教授惠赠．SB液体培养基：30gtmptone,20g酵母膏,10gMops溶于IL去离子水中,调pH7．0．枝…

Construction of Random Peptide Libraries for Screening the Motif of Anti-virus Antibodies

A "fusion phase" is a filamentous virion displaying on its surface a foreign peptide fused to a coat protein (pⅢ or pⅧ),and harboring the gene for the fusion protein within its genome.Important biotech achievements include identification of peptide molecules that mimic the structure and function of sugars and carbohydrates that are difficult to synthesize, peptides that are effective vaccines. A 78-bases template encoding twelve peptides and biothinylated primers in 5' terminus were designed. After PCR amplification, digestion with XhoⅠ/SpeⅠ, ligation and electrotransfection, a 5 × 10 7 clone libraly was estabished. Next,screening anti-idiotype peptides by antiHIV antibodies from the peptide library is expected.