pComb3

INTRODUCTION

pComb3 is a phage display plasmid containing f1 ori and geneIII sequence from M13 filamentous phage. By transformation of pComb3 into XLI-Blue cells, with the help of VCSM13 phages, phages displaying protein of interest can be produced, panned and screened for positive clones.

pComb3 plasmid contains two expression cassettes: the SacI/XbaI restriction sites for cloning the light chain and the XhoI/SpeI restriction sites for cloning the heavy chain. Both the heavy chain and the light chain are N-terminal tagged with pelB leader peptide directing the fusion proteins to the periplasmic space of Gram-negative bacteria where pelB is removed and Fab is presented on the surface of the intact phages. M13 geneIII encoding sequence can be excised using SpeI/NheI double enzyme digestion to allow soluble Fab expression.

pComb3 was first designed for Fab phage displlay. However, besides Fab, pComb3 vector system also can be used for displaying other proteins of interest.

VECTOR INFORMATION

Figure 1. pComb3 Vector Map

Figure 2. NcoI enzyme digestion of pComb3 (1013 bp + 3016 bp)

RESEARCH REFERENCES

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