Shotgun phage display cloning.
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 phage mid-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% 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%.
Department of Microbiology, Swedish University of Agricultural Sciences, Box 7025, UPPSALA, SE-750 07 UPPSALA, Sweden. Karin.Jacobsson@mikrob.slu.se
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D001435:Bacteriophages / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D020650:Combinatorial Chemistry Techniques; D011993:Recombinant Fusion Proteins / Q000235:genetics*
D011993:Recombinant Fusion Proteins