Production of bivalent protein EspA-Tir from Escherichia coli O157:H7 in Tobacco (Nicotiana tobbacum)

E. coli O157: H7 is an important zoonotic pathogen that causing severe gastrointestinal disease such as hemorrhagic diarrhea and hemolytic uremic syndrome. The first step of bacterial pathogenicity is، attaching to the host cell. EspA is a protein molecule and forms as filamentous structure that is transiently expressed on the outer membrane surface and interacts with the host cell during the early stage adhesion and forms bacterial biofilm، this filamentous structure make it a strong immunogenic. Tir is bacterial protein that translocated to host cell after expression in the bacteria by type III secretion system (T3SS) and integrated in to host cell membrane. Intimin can attach to the host cell by conducting to Tir. The purpose of this study is constructing bivalent gen which contains virulence factor mention before and transferring in to target plant in order to production edible vaccine against E. coli O157: H7. After bioinformatics investigation the two bivalent gen construction (espA-Tir،) were attached by peptide linker، and the gens were constructed. After codon optimization based on tobacco; construction were cloned in expression vector which contains CaMV35s promoter. After transformation and regeneration، the expressions of transgenes were showed in analysis.