Cloning and expression of the V-domain of the CD166 in prokaryotic host cell
Purpose: CD166/ALCAM (Activated leukocyte cell adhesion molecule) as an immunoglobulin is implicated in cell migration. It is also involved in tumorigenesis of CRC (colorectal cancer) and known as a cancer stem cell marker. CD166, as a membrane protein, potentially represents either diagnostic or therapeutic capacities for CRC.
Methods: In this study, the sequence of V domain was optimized for expression in prokaryotic host using online tools and cloned into pET-28a plasmid. The recombinant pET28a was transformed into the E. coli BL21DE3 using heat shock method and expression of recombinant V domain was examined using SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis).
Results: The results confirmed protein expression of recombinant 22.77 kDa V domains in bacterial expression system.
Conclusion: V domain of the CD166 was expressed successfully in E. coli bacteria. This recombinant fragment can be introduced as a suitable diagnostic and therapeutic candidate for screening and cancer-therapy of CRC patients, respectively.
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