制备 VCSM13 辅助噬菌体，用于显示文库重扩增和生物平移。

Cold Spring Harbor protocols Pub Date : 2024-08-16 DOI:10.1101/pdb.prot108569

Hyunji Yang, Jisu Chae, Hyori Kim, Jinsung Noh, Junho Chung

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引用次数: 0

摘要

噬菌体展示抗体文库可以使用任何易于免疫的物种来构建。pComb3XSS 噬菌体载体常用于文库克隆和噬菌体展示。这种噬菌体编码丝状噬菌体 f1 的复制源，但缺乏复制和组装噬菌体颗粒所需的所有基因。因此，从这些噬菌体中复制和组装噬菌体需要一种 "辅助 "噬菌体，这种噬菌体能提供文库生产和生物筛选过程中这些步骤所必需的基因。VCSM13 就是辅助噬菌体之一。在本方案中，我们将介绍 VCSM13 辅助噬菌体的制备方法。用户应准备 VCSM13 辅助噬菌体用于文库再扩增和生物筛选。

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Preparation of VCSM13 Helper Phage for Display Library Reamplification and Bio-Panning.

Phage-displayed antibody libraries can be constructed using any species that is easily immunized. The pComb3XSS phagemid vector is commonly used for library cloning and phage display. This phagemid encodes the origin of replication of the filamentous bacteriophage f1 but lacks all the genes required for replication and assembly of phage particles. The replication and the assembly of phage from these phagemids thus requires a "helper" phage that provides the genes essential for those steps during library production and bio-panning. One of those helper phages is VCSM13. In this protocol, we describe the preparation of VCSM13 helper phage. Users should prepare VCSM13 helper phage for library reamplification and for bio-panning.

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来源期刊

Cold Spring Harbor protocols Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)

CiteScore

3.00

自引率

0.00%

发文量

163

期刊介绍： Cold Spring Harbor Laboratory is renowned for its teaching of biomedical research techniques. For decades, participants in its celebrated, hands-on courses and users of its laboratory manuals have gained access to the most authoritative and reliable methods in molecular and cellular biology. Now that access has moved online. Cold Spring Harbor Protocols is an interdisciplinary journal providing a definitive source of research methods in cell, developmental and molecular biology, genetics, bioinformatics, protein science, computational biology, immunology, neuroscience and imaging. Each monthly issue details multiple essential methods—a mix of cutting-edge and well-established techniques.