Lambdoid bacteriophage vectors for expression and display of foreign proteins
AssigneeThe Scripps Research Institute
InventorsIchiro Maruyama, Hiroko Maruyama, Sydney Brenner
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Abstract
Lambdoid phage comprising a matrix of proteins encapsulating a genome encoding first and second polypeptides of an autogenously assembling receptor and a receptor comprised of the first and second polypeptides surface-integrated into the matrix via a lambdoid phage tail protein matrix anchor domain fused to at least one of the polypeptides.
Claims22
A recombinant lambdoid bacteriophage vector comprising a nucleotide sequence that (i) defines the lambdoid elements for replication and packaging of the vector into an assembled bacteriophage, and (ii) encodes a conditionally suppressible cistron for expression of a tail protein and a fusion protein that comprises:
- a) a promoter for transcribing the cistron,
- b) a first upstream translatable sequence that encodes a pV lambdoid bacteriophage tail polypeptide,
- c) a first ribosome binding site to initiate translation of said upstream translatable sequence,
- d) a second translatable sequence operatively linked downstream to said first translatable sequence that (1) encodes a linker polypeptide in frame with said tail polypeptide and (2) includes a sequence adapted for ligation of an insert polynucleotide that defines a third translatable sequence downstream from said second translatable sequence that encodes a preselected polypeptide, and
- e) a suppressor termination codon within said second translatable sequence that upon suppression results in read-through to form a fusion polypeptide consisting of said tail polypeptide, linker polypeptide and preselected polypeptide.
A recombinant lambdoid bacteriophage comprising a matrix of proteins encapsulating a lambdoid genome encoding a fusion protein, said matrix including said fusion protein, wherein the fusion protein is surface accessible in said matrix, and said fusion protein comprising, in the direction of amino terminus to carboxy terminus, a pV lambdoid bacteriophage tail polypeptide, a linker polypeptide and a preselected polypeptide.
A library of recombinant lambdoid bacteriophage particles wherein each particle comprises a matrix of proteins encapsulating a lambdoid genome, said matrix including a fusion protein having an amino acid residue sequence that comprises, in the direction of amino terminus to carboxy terminus, a pV lambdoid bacteriophage tail polypeptide, a linker polypeptide and a preselected polypeptide defining a biological activity, wherein said fusion protein is surface accessible in said matrix.
Dependencies are inferred from the claim text.