Shotgun Phage Display Cloning

Authors: Jacobsson K.; Frykberg L.

Source: Combinatorial Chemistry & High Throughput Screening, Volume 4, Number 2, April 2001 , pp. 135-143(9)

Publisher: Bentham Science Publishers

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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

Language: English

Document Type: Review article

DOI: 10.2174/1386207013331255

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