Molecular Cloning and Expression of α-Amylase Gene of Human Pancreatic Amylase (Amy2a) in Escherichia Coli

Abstract

α-Amylase is an enzyme that acts as a catalyst for the hydrolysis of alpha-linked polysaccharides into α-anomeric products(1). The enzyme can be derived from a variety of sources, each with different characteristics. α-Amylase found within the human body serves as the enzyme active in pancreatic juice and salvia(2).α-Amylase is not only essential in human physiology but has a number of important biotechnological functions in various processing industries. the aim of this study is cloning and expression of α-Amylase gene of Human pancreatic amylase (amy2A)in Escherichia coli.to achieve this aim a gene coding for α-Amylase from NCBI was selected and amplified with specific primers and PCR.PCR product was cloned into a bacterial expression vector pET21-a . The recombi- nant plasmid was used to transform the bacterial expression strain BL21 and expression products were analyzed with SDS-PAGE, and Immunoblotting. Based on these results, the bacterial expression system successfully ex- pressed α-Amylase.