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.
summary
Bacterial Resistance(s): Ampicillin
Growth Temperature: 37°C
Growth Strain(s): TG1
Copy number: High Copy
Insert Size (bp): 1600
Promoter:lacZ
Tags / Fusion Proteins
geneVIII (C terminal on backbone)