AMPLIFICATION, CLONING AND EXPRESSION OF THE REG3 Δ GENE FROM MOUSE PANCREAS

Abstract

Introduction: Reg proteins are a group of regenerating proteins which are implicated in the pancreas
developmental biology. The aim of the study was to explore suitable conditions for cloning and expression of
this recombinant protein that can ultimately be explored in further studies for testing its potential as a promising
therapeutic for diabetes by inducing regeneration/neogenesis of pancreatic beta cells. Methodology: Total RNA
was isolated from BALB/C mouse pancreas, cDNA was synthesized and Reg3 δ gene was amplified with gene
specific primers. A recombinant plasmid (pET28a vector) was constructed with Reg3 δ gene. Recombinant
protein was expressed in BL21 (DE3) strain in LB media. Expressed protein was isolated and separated on 12%
SDS-PAGE. Results: Clones were confirmed with restriction and colony PCR and SDS-PAGE confirmed the 16
kDa band in IPTG induced samples. Further work for the purification of this protein will be pursued in future.