Phagemid vectors

Abstract

Phagemid vectors containing a sequence of features between a Col E1 origin and an f1 origin are useful for display of polypeptides or proteins, including antibody libraries.

1. A phagemid vector comprising:
 a selectable marker;    a ColE1 origin;    an f1 origin; and    after the ColE1 origin but before the f1 origin, further comprising the following features:    a bacterial transcription terminator;    a promoter,    a first ribosomal binding site;    a first leader sequence;    a first cloning region;    a second ribosomal binding site;    a second leader sequence;    a second cloning region for receiving a gene encoding a polypeptide to be displayed; and    a nucleotide sequence encoding a product that enables display of a polypeptide on the surface of a phagemid particle.    
     
     
       2. A phagemid vector as in  claim 1  wherein at least one of the first or second ribosomal binding sites comprises Seq. ID No. 13. 
     
     
       3. A phagemid vector as in  claim 1   A phagemid vector comprising:
   a selectable marker;    
   a ColE 1  origin;    
   an f 1  origin; and    
   after the ColE 1  origin but before the f 1  origin, further comprising the following features: a bacterial transcription terminator and a promoter wherein the bacterial transcription terminator is upstream of the promoter;      a first ribosomal binding site;        a first leader sequence;        a first cloning region;        a second ribosomal binding site;        a second leader sequence;        a second cloning region for receiving a gene encoding a polypeptide to be displayed; and        a nucleotide sequence encoding a product that enables display of a polypeptide on the surface of a phagemid particle,      
 wherein at least one of the first or second leader sequences comprises a sequence selected from the group consisting of Seq. ID No. 14 and Seq. ID No. 17.  
 
     
     
       4. A phagemid vector as in  claim 1  wherein the nucleotide sequence encoding a product encodes a protein selected from the group consisting of pIII and pVIII. 
     
     
       5. A phagemid vector as in  claim 1  wherein the nucleotide sequence encoding a product encodes a truncated pIII. 
     
     
       6. A phagemid vector as in  claim 1  wherein the nucleotide sequence encoding a product encodes a synthetic pIII. 
     
     
       7. A phagemid vector as in  claim 1  wherein the selectable marker is selected from the group consisting of ampicillin resistance, chloramphenicol transferase resistance, tetracycline resistance and kanamycin resistance. 
     
     
       8. A phagemid vector comprising Seq. ID No. 18. 
     
     
       9. A vector comprising a sequence selected from the group consisting of Seq. ID Nos. 19, 20 and 21.