Adaptor-directed helper systems

• US 20060281181A1
• Filed: 12/19/2005
• Published: 12/14/2006
• Est. Priority Date: 11/02/2001
• Status: Active Grant

• Current Assignee: Caili Wang, Pingyu Zhong, Xinwei Wang
• Original Assignee: Caili Wang, Pingyu Zhong, Xinwei Wang
• Inventors: Zhong, Pingyu, Wang, Caili, Wang, Xinwei
• Granted Patent: US 7,910,350 B2
• Time in Patent Office: Days
• US Class Current: 435/457
• CPC Class Codes: C12N 15/1037 Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display
• CPC Class Codes: C40B 40/02 Libraries contained in or displayed by microorganisms, e.g. bacteria or animal cells; Libraries contained in or displayed by vectors, e.g. plasmids; Libraries containing only microorganisms or vectors

Abstract

The present invention provides adapter-directed display systems for expressing exogenous polypeptide within a host cell and/or displaying the exogenous polypeptide on the outer surface of a genetic package. This subject systems are particularly useful for displaying a genetically diverse repertoire of monomeric and multimeric polypeptides. The invention also provides both expression and helper vectors and kits containing components of the subject display systems. Also provided are genetic packages displaying the exogenous polypeptides of particular interest. Further provided by the invention are methods of using the subject display systems.

Claims

1-20. -20. (canceled)

21. A helper vector for displaying a polypeptide on the outer surface of a genetic package comprising:

outer-surface sequences necessary for packaging the genetic package, wherein at least one of the surface presenting sequences is fused in-frame to an adapter, said adapter acting, when the polypeptide is produced in a suitable host cell, to cause the display of the polypeptide.

22. The helper vector of claim 21, wherein the vector is a phage helper vector.

23. The helper vector of claim 21, wherein the vector is a bacterial helper vector.

24. The helper vector of claim 21, wherein the genetic package is selected from the group consisting of viruses, cells, and spores.

25. The helper vector of claim 22, wherein the outer-surface sequences encode functional coat proteins of a phage.

26. The helper vector of claim 22, wherein the phage is filamentous phage.

27. The helper vector of claim 22, wherein in the outer-surface sequences are selected from the group consisting of gene III, gene VI, gene VII, gene VIII, and gene IX of a filamentous phage.

28. The helper vector of claim 23, wherein the outer-surface sequences encode bacterial outer-surface proteins.

29. The helper vector of claim 23, wherein the bacterial outer-surface proteins are selected from the group consisting of Lpp-OmpA, TraT, Pal, Opr1, Inp and AIDA-I.

30. The helper vector of claim 21, wherein the adapter causes the display of the polypeptide via pairwise interaction with a second adapter, which is fused in-frame with the polypeptide.

31. The helper vector of claim 21, wherein the adapter causes the display of the polypeptide in the absence of expression of an outer-surface protein via a phagemid vector or a plasmid.

32. The helper vector of claim 21, wherein the two adapters are heterodimerization sequences.

33. The helper vector of claim 21, wherein the two adapters are homodimerization sequences.

34. The helper vector of claim 21, wherein the two adapters are homodimerization sequences.

35. The helper vector of claim 34, wherein the homodimerization sequences consist of a pair of cysteine residues.

36. The helper vector of claim 32, wherein the two adapters form a coiled-coil dimer.

37. The helper vector of claim 36, wherein the two adapters are leucine zippers.

38. The helper vector of claim 36, wherein the two adapters comprise heterodimeric receptor sequences that mediate heterodimerization of the receptors.

39. The helper vector of claim 36, wherein the two adapters comprise heterodimerization sequences of GABA_B receptor 1 and GABA_B receptor 2, respectively.

40. The helper vector of claim 36, wherein the two adapters comprise heterodimerization sequences of GABA_B receptor 2 and GABA_B receptor 1, respectively.

41-76. -76. (canceled)