dual promoter vector construction for simultaneous gene expression using spliced overlap extension by polymerase chain reaction (soe-pcr) technique

dual promoter vector construction for simultaneous gene expression using spliced overlap extension by polymerase chain reaction (soe-pcr) technique

m.h. sekhavati, m. tahmoorespur, t. abbassi-daloii, s. yousefi, a.a. khabiri

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.