pComb8
(Plasmid #63889)

• Purpose: (Empty Backbone) For phage display of antibody libraries by fusion to modified geneVIII.
• 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:
  • geneVIII (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

pComb8 is nearly identical to the original pComb3 vector but contains the phage pVIII fusion protein for multi-valent expression along the sides of the phage via fusion with pVIII. It 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-geneVIII fusion product assemble in the periplasm to form the Fab fragment. Soluble Fab can be expressed by removing the gene for the pVIII phage fusion protein by SpeI/NheI digest. Sufficient soluble protein is also found in the periplasmic space as a result of proteolysis.