Recombinant Production of a Chimeric Protein Containing StxB, Eae, Cfab, and LTB Proteins in Hairy Roots of Nicotiana tabaccum

Document Type : Original Article


1 Department of Agricultural Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran

2 Applied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran


Introduction: The technology of recombinant protein production has revolutionized biological, pharmaceutical, and medical researches. Plants are appropriate systems for the production of recombinant proteins. In the present study, a chimeric gene encoding a chimeric protein containing StxB, Eae, CfaB, and LTB as a candidate vaccine against enterotoxigenic and enterohemorrhagic E. coli, was transferred to the hairy root of the tobacco plant and its expression was analyzed.
Materials and Methods: For this aim, the plant-based codon-optimized gene, which was synthetically cloned in pBI121 binary vector, was transferred into Agrobacterium rhizogenes competent cells. A. rhizogenes cells containing pBI121+secl were transferred into sterile Nicotiana tabaccum plants by direct and indirect methods. Finally, the expression of recombinant protein was analyzed by the ELISA method.
Results: The transformation of A. rhizogenes cells and Tobacco plants was confirmed by PCR amplification of a fragment of the chimeric gene with specific primers. SDS-PAGE and ELISA analysis showed the successful expression of the chimeric recombinant protein.
Conclusions: In conclusion, it was shown that the hairy roots of tobacco plant is a proper expression system for the recombinant production of SECL protein. The protein can be used for mice's immunization against enterotoxigenic and enterohemorrhagic E. coli strains.