Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Patent
1995-02-22
1998-06-23
Guzo, David
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
435 6, 435 71, 4351723, 43525233, 4353201, C12Q 168, C12Q 170, C12N 121, C12N 1564
Patent
active
057703566
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to cloning vectors and methods for producing a library of DNA molecules capable of expressing a heterodimeric receptor and a second dimeric heterologous indicator fusion polypeptide on the surface of a filamentous phage particle.
BACKGROUND
Filamentous bacteriophages are a group of related viruses that infect bacteria. They are termed filamentous because they are long and thin particles comprised of an elongated capsule that envelopes the deoxyribonucleic acid (DNA) that forms the bacteriophage genome. The F pili filamentous bacteriophage (Ff phage) infect only gram-negative bacteria by specifically adsorbing to the tip of F pili, and include fd, f1 and M13.
The mature capsule of Ff phage is comprised of a coat of five phage-encoded gene products: cpVIII, the major coat protein product of gene VIII that forms the bulk of the capsule; and four minor coat proteins, cpIII and cpIV at one end of the capsule and cpVII and cpIX at the other end of the capsule. The length of the capsule is formed by 2500 to 3000 copies of cpVIII in an ordered helix array that forms the characteristic filament structure. About five copies each of the minor coat proteins are present at the ends of the capsule. The gene III-encoded protein (cpIII) is typically present in 4 to 6 copies at one end of the capsule and serves as the receptor for binding of the phage to its bacterial host in the initial phase of infection. For detailed reviews of Ff phage structure, see Rasched et al., Microbiol. Rev., 50:401-427 (1986); and Model et al., in "The Bacteriophages, Volume 2", R. Calendar, Ed., Plenum Press, pp. 375-456 (1988).
The assembly of a Ff phage particle involves highly complex mechanics. No phage particles are assembled within a host cell; rather, they are assembled during extrusion of the viral genome through the host cell's membrane. Prior to extrusion, the major coat protein cpVIII and the minor coat protein cpIII are synthesized and transported to the host cell's membrane. Both cpVIII and cpIII are anchored in the host cell membrane prior to their incorporation into the mature particle. In addition, the viral genome is produced and coated with cpV protein. During the extrusion process, cpV-coated genomic DNA is stripped of the cpV coat and simultaneously re-coated with the mature coat proteins. The assembly mechanisms that control transferral of these proteins from the membrane to the particle is not presently known.
Both cpIII and cpVIII proteins include two domains that provide signals for assembly of the mature phage particle. The first domain is a secretion signal that directs the newly synthesized protein to the host cell membrane. The secretion signal is located at the amino terminus of the polypeptide and targets the polypeptide at least to the cell membrane. The second domain is a membrane anchor domain that provides signals for association with the host cell membrane and for association with the phage particle during assembly. This second signal for both cpVIII and cpIII comprises at least a hydrophobic region for spanning the membrane.
cpVIII has been extensively studied as a model membrane protein because it can integrate into lipid bilayers such as the cell membrane in an asymmetric orientation with the acidic amino terminus toward the outside and the basic carboxy terminus toward the 100 inside of the membrane. The mature protein is about 50 amino acid residues in length of which 11 residues provide the carboxy terminus, 19 residues provide the hydrophobic transmembrane region, and the remaining residues comprise the amino terminus. Considerable research has been done on the secretion signal region of cpVIII to advance the study of membrane protein synthesis and targeting to membranes. However, little is known about the changes that are tolerated in the structure of the cpVIII membrane anchor region that would allow for assembly of phage particles.
Manipulation of the sequence of cpIII shows that the C-terminal 23 amino acid residue stretch of hydrophobic amino acids
REFERENCES:
patent: 5223409 (1993-06-01), Ladner
patent: 5403848 (1995-04-01), Ladner
Jaenicke, Prog. Biophys. Malec. Biol., vol. 49, pp. 117-237, 1987.
Bass, et al., "Hormone Phage: An Enrichment Method for Variant Proteins With Altered Binding Properties", Proteins: Structure, Function and Genetics, 8:309-314 (1990).
Bird, et al., "Single-Chain Antigen-Binding Proteins", Science, 242:423-6 (1988).
Boss, et al., "Assembly of Functional Antibodies from Immunoglobulin Heavy and Light Chains Synthesized in E. coli", Nuc. Acids Res., 12:3791-3806(1984).
Cabilly, et al., Generation of Antibody-Activity from Immunoglobulin Polypeptide Chains Produced in Escherichia coli, Proc. Natl. Acad. Sci., USA 81:3273-3277 (1984).
Caton, et al., "Influenza Virus Hemagglutinin-Specific Antibodies Isolated From a Combinatorial Expression Library are Closely Related to the Immune Response of the Donor", Proc. Natl. Acad. Sci., USA, 87:6450-6454 (1990).
Devlin, et al., "Random Peptide Libraries: A Source of Specific Protein Binding Molecules", Science, 249:404-406 (1990).
Larrick, et al., "Polymerase Chain Reaction Using Mixed Primers: Cloning of Human Monoclonal Antibody Variable Region Genes from Single Hybridoma Cells", Biotechnology, 7:934-938(1989).
Marks, et al., "By-Passing Immunization Human Antibodies from V-Gene Libraries Displayed on Phage", J. Mol. Biol., 222:581-597 (1991).
Model, et al., "Filamentous Bacteriophage", ed. R. Calendar, Plenum Publishing Corp., 2:375-456 (1988).
Mullinax, et al., "Identification of Human Antibody Fragment Clones Specific for Tetanus Toxoid in a Bacteriophage Alpha Immunoexpression Library", Proc. Natl. Acad. Sci., USA, 87:8095-8099 (1990).
O'Neill, et al., "Design of DNA-Binding Peptides Based on the Leucine Zipper Motif", Science, 249:774-778 (1990).
Orlandi, et al., "Cloning Immunoglobulin Variable Domains for Expression by the Polymerase Chain Reaction", Proc. Natl. Acad. Sci., USA, 86:3833-3837 (1989).
Pluckthun, A., "Antibodies from Escherichia Coli", Nature, 347:497-498 (1990).
Rasched, et al., "Ff Coliphages: Structural and Functional Relationships", Microbiol. Rev., 50:401-427 (1986).
Roberts, et al., "Protease Inhibitor Display M13 Phage: Selection of High-Affinity Neutrophil Elastase Inhibitors", Gene, 121:9-15 (1992).
Roberts, et al., "Directed Evolution of a Protein: Selection of Potent Neutrophil Elastase Inhibitors Displayed on M13 Fusion Phage", Proc. Natl. Acad. Sci., USA, 89:2429-2433 (1992).
Sastry, et al., "Cloning of the Immunological Repertoire in E. coli for Generationof Monoclonal Catalytic Antibodies: Construction of a Heavy Chain Variable Region-Specific cDNA Library", Proc. Natl. Acad. Sci., USA, 86: 5728-5732 (1989).
Scott, et al., "Searching for Peptide Ligands with an Epitope Library", Science, 249:386-390 (1990).
Short, et al.,Lambda ZAP: A Bacteriophage Lambda Expression Vector with in vitro Excision Properties, Nuc. Acids Res., 16:7583-7600 (1988).
Skerra, et al., "Assembly of a Functional Immunoglobulin Fv Fragment in E. coli" Science, 240:1038-1043 (1988).
Skerra, et al., "The Functional Expression of Antibody Fv Fragments in E. coli Improved Vectors and a Generally Applicable Purification Technique", Biotechnology, 9:273-278 (1991).
Smith, George P., "Filamentous Fusion Phage: Novel Expression Vectors That Display Cloned Antigens on the Virion Surface", Science, 228:1315-1317 (1985).
Solazzo, et al., "Expression of an Exogenous Peptide Epitope Genetically Engineered in the Variable Domain of an Immunoglobulin: Implications for Antibody and Peptide Folding", Protein Engineering, 4:215-220 (1990).
Takeda, et al., Construction of Chimaeric Processed Immunoglobulin Genes Containing Mouse Variable and Human Constant Region Sequences, Nature, 314:452-454 (1985).
Ward, et al., "Binding Activities of a Repertoire of Single Immunoglobulin Variable Domains Secreted from Escherichia coli", Nature, 341:544-546 (1989).
Barbas, et al., Methods: A Companion to Methods in Enzymology, 2: 119-124 (1991).
Barbas, et a
Lerner Richard A.
Light, II James Paul
Fitting Thomas
Guzo David
Holmes Emily
The Scripps Research Institute
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