Dual Promoter Vector Construction for Simultaneous Gene Expression Using Spliced Overlap Extension by Polymerase Chain Reaction (SOE-PCR) Technique

Abstract

There are two different co-expression systems including bicistronic; dual-vector or two-promoter to express two different genes simultaneously and also to study protein-protein interactions. Bicistronic system has disadvantages e.g. compared with two-promoter system. In this paper, a simple method based on spliced overlap extension by polymerase chain reaction (SOE-PCR) technique was demonstrated for construction of two-promoter vector to express two genes equally. To construct two-promoter vector, two pairs of mega-primer containing enzymatic restriction sites, T7 promoter, Lac operator and ribosome binding site (RBS) sequences were used in SOE-PCR. The constructed vector can be used in order to co-expression of other genes properly in a variety of bacterial expression hosts.