Assembly of functional bacteriophage lambda virions incorporating C-terminal peptide or protein fusions with the major tail protein.
The tolerance of bacteriophage lambda morphogenesis for C-terminal additions to the tail tube major protein subunit (the V gene product; gpV) has been investigated. A second modified copy of the lambda V gene, either within a novel phage vector itself or plasmid-borne, was expressed during phage growth. High-level substitution of wild-type gpV by modified gpV bearing a basic C-terminal peptide sequence (RRASV; a target site for cAMP-dependent protein kinase) was possible using multiple repeats of a serine-glycine (SGGG) linker sequence. Highly purified phage bearing copies of gpV-RRASV could be efficiently phosphorylated by the appropriate protein kinase, and the incorporated label was shown to migrate exclusively at the expected size in protein gels. A large tetrameric protein (beta-galactosidase) could be incorporated into active virions in at least one copy, again using a Ser-Gly linker. These studies suggest that with a suitable spacing linker and controlled levels of expression, it is likely that a wide range of protein or peptide substitutents can be fused with gpV at its C terminus and assembled as component subunits of the tail tube.
Dunn
I S
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Department of Pathology, Medical School University of Queensland Brisbane, Australia.
D000595:Amino Acid Sequence; D010582:Bacteriophage lambda / Q000235:genetics* / Q000502:physiology / Q000648:ultrastructure; D001483:Base Sequence; D004279:DNA, Viral; D005814:Genes, Viral / Q000235:genetics; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D015678:Viral Structural Proteins / Q000235:genetics; D017376:Viral Tail Proteins / Q000235:genetics* / Q000502:physiology; D014771:Virion / Q000235:genetics / Q000502:physiology; D014779:Virus Replication; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D004279:DNA, Viral; D011993:Recombinant Fusion Proteins; D015678:Viral Structural Proteins; D017376:Viral Tail Proteins; D001616:beta-Galactosidase