Extending filamentous phage host range by the grafting of a heterologous receptor binding domain.
fd and IKe are two similar filamentous phage which infect their hosts by means of pili found on the host membrane: fd infects bacteria bearing F pili, whereas IKe infects bacteria bearing N or I pili. Infection is mediated by the gene 3 protein (g3p), which of the nine proteins found in both phage is the most diverse. Previous attempts to incorporate g3p from one phage into the other by complementation have been unsuccessful [Bross et al. (1988) J. Gen. Microbiol. 134, 461-471]. Here we have grafted different parts of IKe g3p to the end of fd g3p and so augmented the host range of fd phage. We show that phage bearing such chimeric g3p are able to infect bacteria bearing both N and F pili providing they contain at least the receptor domain of IKe g3p, the infection of N bearing bacteria occurring at a level 70,000 times greater than background. This level of infection can be increased tenfold by including the glycine-rich domain as well. Addition of the penetration domain does not improve the level of infection above that of the receptor domain alone, indicating that the fd penetration domain is functional in the infection of bacteria bearing either N or F pili. Similarly derived fd phagemid also show increased infection of bacteria bearing N pili, albeit at much lower levels, suggesting that efficient infection requires more than one functional g3p on the surface of the phage.
Marzari
R
R
International School for Advanced Studies (SISSA), Trieste, Italy.
Sblattero
D
D
Righi
M
M
Bradbury
A
A
D001419:Bacteria / Q000737:chemistry / Q000821:virology; D001665:Binding Sites / Q000235:genetics*; D015153:Blotting, Western; D036022:Capsid Proteins; D002678:Chimera / Q000235:genetics*; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000235:genetics*; D010861:Fimbriae, Bacterial / Q000737:chemistry / Q000235:genetics / Q000502:physiology; D017912:Inovirus / Q000235:genetics*; D011485:Protein Binding / Q000235:genetics*; D014760:Viral Fusion Proteins / Q000235:genetics*
D036022:Capsid Proteins; D004268:DNA-Binding Proteins; D014760:Viral Fusion Proteins