Phage fusion

Lindner P, Bauer K, Krebber A, Nieba L, Kremmer E, Krebber C, Honegger A, Klinger B, Mocikat R, Pluckthun A, Specific detection of his-tagged proteins with recombinant anti-His tag scFv-phosphatase or scFv-phage fusions, Bio techniques, 22 140-149, 1997. 

Smith, G.P. Filamentous fusion phage novel expression vectors that display cloned antigens on the virion surface. Science 228 1315-1317, 1985. 

The advantage of the Jun-Fos phage display system is that the fusion of the library DNA is to the C-terminus of a protein rather than between the... 

The pJuFo system also has the potential to display homodimers or homomultimeric proteins on the surface of the phage. This is possible because the protein of interest is expressed independent of the phage proteins. Therefore, the Fos-ORF fusion protein is secreted to the periplasm, the Fos portion interacts with Jun to attach the protein to the phage, and other copies of the Fos-ORF protein can interact with each other to form a dimer or multimer. However, the successful display of homodimers has not been demonstrated in practice. 

A variation on the theme has been to map out protease specificity.28 A library of fusion proteins was constructed in a modular manner. The synthetic protein had an N-terminal domain that binds very tightly to an affinity column. This domain was connected to the C-terminal domain of M13 gene III by a randomized peptide sequence. The phages were then bound to the affinity support and treated with a protease. Phages that had a protease-susceptible site were cleaved from the support and eluted. This procedure was subsequently used to map out the specificity of furin,29 which is described in the next chapter. 

A number of phagemid display systems that fuse Ab chains to either full-length or N-terminally deleted gp3 fusions have been described, along with the polymerase chain reaction (PCR) primer sets, restriction enzymes, and host strains required for the display of scFvs and Fabs (3—7). Some systems are commercially available. There is an important difference in whether the Ab-gp3 fusions are with the whole of the protein or with just the C-terminal domain. Fusions to the whole of gp3 require that the expression of the fusion is suppressed until after it is infected with the helper phage because of the resistance... 

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