pComb3
(Plasmid #63888)

• Purpose: (Empty Backbone) For phage display of antibody libraries by fusion to modified geneIII.
• Depositing Lab: Carlos Barbas
• Publication: Barbas CF et al Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):7978-82.

Backbone

• Vector backbone: pBluescript
• Backbone size (bp): 3000
• Vector type: Bacterial Expression ; phage display
• Promoter: lacZ
• Tag / Fusion Protein:
  • geneIII (C terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: High Copy

Cloning Information

• Cloning method: Restriction Enzyme

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The original pComb3 vector was designed for phage display of Fabs which are cloned into two separate cloning cassettes one chain at a time using SacI/XbaI restriction sites (for the light chain) and XhoI/SpeI restriction sites (for the heavy chain). The heavy chain is expressed fused to the pVIII. Both cassettes have the pelB leader sequence, the light chain and heavy chain-geneIII fusion product assemble in the periplasm to form the Fab fragment. This vector system has also been used for the display of a wide variety of other proteins like zinc fingers, peptides, and cDNA fragments. Phage can be produced which express Fab or other proteins or peptides of interest fused to the phage pIII protein for expression on the head of the phage. Soluble Fab can be expressed by removing the gene for the pIII phage fusion protein by SpeI/NheI digest. Sufficient soluble protein is also found in the periplasmic space as a result of proteolysis.