Patent ID: 7410801

Claim:
A process of obtaining recombinant lambdoid bacteriophage with high density display of functional peptides and proteins on surface of said bacteriophage comprising the steps of: a) Constructing a donor plasmid comprising: A first recombination sequence, and a second recombination sequence, wherein said first and second recombination sequences are incompatible with each other, and wherein said first and second recombination sequences flank a nucleotide sequence that defines the elements for replication of the vector in bacteria, a nucleotide sequence encoding a first selectable marker, and a first inducible cistron comprising: a promoter for transcribing said first cistron, a first ribosome binding site, a first translatable nucleotide sequence encoding a lambdoid bacteriophage capsid polypeptide a second translatable sequence operatively linked to said first translatable sequence and encoding a first linker polypeptide, a protease cleavable peptide sequence, a second linker polypeptide, and a sequence adapted for ligation of an insert polynucleotide or an MCS; wherein each translatable nucleotide sequence is in the same reading frame such that induction of said first cistron expresses a fusion polypeptide consisting of said capsid polypeptide, linker polypeptide, protease cleavage site, linker polypeptide, and said MCS-encoded or said insert-encoded polypeptide; nucleotide sequence comprising: lambdoid replication and packaging elements, said first recombination sequence, and said second recombination sequence, wherein said first and second recombination sequences flank a second inducible cistron for expression of a second selectable marker transferring the polynucleotide sequences located between said first and second recombination sequences of said donor plasmid to said recipient phage to obtain cointegrates; d) propagating said cointegrates in liquid medium selective for said first selectable marker; and e) harvesting phages displaying said fusion polypeptide.