Dual Promoter Vector Construction for Simultaneous Gene Expression Using Spliced Overlap Extension by Polymerase Chain Reaction (SOE-PCR) Technique
Subject Areas : Camelم.ه. سخاوتی 1 , م. طهمورثپور 2 , ط. عباسی-دلوئی 3 , س. یوسفی 4 , ع.ا. خبیری 5 , ر. اکبری 6 , E. شکاری 7
1 - Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
2 - Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
3 - Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
4 - Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
5 - Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
6 - Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
7 - Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
Keywords: co-expression, protein-protein interactions, SOE-PCR, two-promoter vector,
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.
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