Narges Nazifi, Mojtaba Tahmoorespur*, Mohammad Hadi Sekhavati*, Alireza Haghparast and Mohammad Ali Behroozikhah Pages 188 - 198 ( 11 )
Background: Using a bacterial expression system such as Escherichia coli (E. coli) is very common for protein expression because of its simplicity, low cost and high efficiency.
Objective: In order to express proteins that contain di-sulfide bands, an oxidative space such as the periplasmic environment of the bacteria is required. Therefore, a leader sequence which named signal peptide (SP) is needed to direct recombinant protein to fold in periplasmic space. Interleukin-2 (IL-2) is a prominent cytokine which known as growth factor for T-cells and typically produced by a variety of immune cells that stimulate and regulate inflammatory and immune responses.
Method: This study was designed to predict the best signal peptides to express IL-2 in E. coli. To predict the best signal peptides for the expression of IL-2 in Gram-negative bacteria (E. coli), forty-five sequences of SPs were extracted from data base. Some most important details such as n, h and c regions of signal peptides and their probability were studied through the signalP software.
Afterwards, physico–chemical features of SPs were analyzed by Portparam and Solpro tools. Finally, secretion-pathway and sub-cellular localization sites were evaluated by PRED-TAT and ProtcompB softwares.
Results: At the end of the in-silico analyzes, it was determined that ccmH, PelB, traU, yohN, lolA, yhcN are the most reliable SPs, respectively, with highest score and best performing to express the IL-2 protein in E. coli.
Signal peptide, IL-2, E. coli, periplasm, interleukin, T-cells, immune cells.
Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Pathobiology Deptartment, School of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Department of Brucellosis, Razi Vaccine and Serum Research institute, agricultural Research Education and Extension Organization (AREEO), Karaj