Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697-4576, USA.
Protein Eng Des Sel. 2012 Apr;25(4):145-51. doi: 10.1093/protein/gzr068. Epub 2012 Jan 27.
Phage display libraries are widely used as tools for identifying, dissecting and optimizing ligands. Development of a simple method to access greater library diversities could expedite and expand the technique. This paper reports progress toward harnessing the naturally occurring diversity generating retroelement used by Bordetella bronchiseptica bacteriophage to alter its tail-fiber protein. Mutagenesis and testing identified four sites amenable to the insertion of <19-residue heterologous peptides within the variable region. Such sites allow auto-generation of peptide libraries surrounded by a scaffold with additional variations. The resultant self-made phage libraries were used successfully for selections targeting anti-FLAG antibody, immobilized metal affinity chromatography microtiter plates and HIV-1 gp41. The reported experiments demonstrate the utility of the major tropism determinant protein of B.bronchiseptica as a natural scaffold for diverse, phage-constructed libraries with heterologous self-made phage libraries.
噬菌体展示文库被广泛用作鉴定、剖析和优化配体的工具。开发一种简单的方法来获得更大的文库多样性，可以加速和扩展该技术。本文报道了利用自然发生的多样性生成逆转录元件的进展，该元件被支气管败血波氏杆菌噬菌体用来改变其尾部纤维蛋白。诱变和测试确定了四个适合在可变区插入 <19 个残基异源肽的位点。这样的位点允许在具有额外变化的支架周围自动生成肽文库。所得的自制噬菌体文库成功地用于针对抗 FLAG 抗体、固定化金属亲和层析微量滴定板和 HIV-1 gp41 的选择。报告的实验证明了 B. bronchiseptica 的主要趋向决定蛋白作为具有异源自制噬菌体文库的多样化、噬菌体构建文库的天然支架的实用性。
