Shotgun Phage Display Cloning

Karin      Jacobsson; Lars      Frykberg

Abstract

Shotgun phage display cloning is a useful tool for studying interactions between bacterial and host proteins. Libraries are constructed by cloning randomly fragmented prokaryotic DNA into phagemid-vectors. Theoretically, these libraries will consist of phages that together display all proteins encoded by the bacterial genome. Selecting a gene III-based library, made from Staphylococcus aureus DNA, against IgG and fibronectin resulted in 20-40 percent positive clones after two pannings. Increasing the number of fusion proteins per phage particle by using gene VIII-based display, increased the frequency of correct clones to 75-100 percent

Keywords: Shotgun, Staphylococcus aureus DNA, Collagen, Elastin, IgG, Fibrinogen, Fibronectin, Transferrin, Phage Display Cloning

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Combinatorial Chemistry & High Throughput Screening

Title: Shotgun Phage Display Cloning

Volume: 4 Issue: 2

Author(s): Karin Jacobsson and Lars Frykberg

Affiliation:

Keywords: Shotgun, Staphylococcus aureus DNA, Collagen, Elastin, IgG, Fibrinogen, Fibronectin, Transferrin, Phage Display Cloning

Abstract: Shotgun phage display cloning is a useful tool for studying interactions between bacterial and host proteins. Libraries are constructed by cloning randomly fragmented prokaryotic DNA into phagemid-vectors. Theoretically, these libraries will consist of phages that together display all proteins encoded by the bacterial genome. Selecting a gene III-based library, made from Staphylococcus aureus DNA, against IgG and fibronectin resulted in 20-40 percent positive clones after two pannings. Increasing the number of fusion proteins per phage particle by using gene VIII-based display, increased the frequency of correct clones to 75-100 percent

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Jacobsson Karin and Frykberg Lars, Shotgun Phage Display Cloning, Combinatorial Chemistry & High Throughput Screening 2001; 4 (2) . https://dx.doi.org/10.2174/1386207013331255

DOI https://dx.doi.org/10.2174/1386207013331255	Print ISSN 1386-2073
Publisher Name Bentham Science Publisher	Online ISSN 1875-5402

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