Purpose(Empty Backbone) For phage display of antibody libraries by fusion to modified geneVIII.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||63889||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
- Backbone size (bp) 3000
Vector typeBacterial Expression ; phage display
- Promoter lacZ
/ Fusion Protein
- geneVIII (C terminal on backbone)
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Growth Strain(s)XL1 Blue
Copy numberHigh Copy
- Cloning method Restriction Enzyme
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
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.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pComb8 was a gift from Carlos Barbas (Addgene plasmid # 63889 ; http://n2t.net/addgene:63889 ; RRID:Addgene_63889)
For your References section:Assembly of combinatorial antibody libraries on phage surfaces: the gene III site. Barbas CF 3rd, Kang AS, Lerner RA, Benkovic SJ. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):7978-82. PubMed 1896445