Evaluation of stability and functional activity of deglycosylated and mutant forms of protective antigen (PA83) of Bacillus anthracis in dried leaves of Nicotiana benthamiana

Anthrax is an acute disease caused by Bacillus anthracis , which affects both people and animals. The production and release of B. Anthracis spores is relatively easy. It is a zoonotic disease originating from Bacillus anthracis, a gram-positive bacterium. There are many different systems for recombinant protein production. Some of these are plant, insect, bacteria, yeast and mammalian cell cultures. The plant expression system has many advantages over other expression systems. The transient gene expression method is used because they have a modification mechanism after eukaryotic translation, and the desired protein can be produced in a very short time. Based on this approach, deglycosylation of PA protective antigen was performed. Dried leaves infiltrated with deglycosylated dPA83- -E and pGR-MQ2 in N. benthamiana were used in this studies. And the properties of obtained the dPA83- -E and pGR-MQ2 deglycosylated mutant, the possibilities of using stabilization and characterization studies in many directions in N. benthamiana plants were also studied. These studies are; the production of the in vivo deglycosylated PA83 and mutated versions in N. benthamiana plant, expression control of their dried samples by western blot and obtaining of dried leaf protein by dissolution and crushing method. SDS-PAGE and WESTERN BLOT analysis of these results were done. With the findings obtained at the end of this study, the evaluation of stabilization, the possibilities of using stabilization and characterization studies in many directions were also studied. When all results are examined, it has been shown that the protein values of dried leaves have been stable for approximately more than 1 year.