فهرست مطالب زهرا مشیری

Saponaria officinalis is a perennial plant of the clove family. Hairy root culture system is a suitable alternative to improve the production of compounds with medicinal value. Extensive application of nanotechnology in hairy root culture is a suitable substrate for the production of active elements. In this study, the effect of different concentrations of titanium dioxide nanoparticles (0, 10, 20, 30 and 50 mg / l) and two exposure times (24 and 48 hours) on growth rate, antioxidant capacity, phenol and flavonoid content, antioxidant enzymes activity, polyphenol compounds content and the amount of SO6 protein in soapy hairy roots were investigated. The highest and lowest fresh weight of hairy roots (3.09 and 0.96 g, respectively) were found in culture medium treated with 10 and 20 mg / l titanium dioxide nanoparticles at 48 and 24 hours of exposure time, respectively. The highest total Phenol (9.79 mg GAE per g FW) and flavonoid (1.06 mg Que per g FW) content were observed in hairy roots treated with 50 and 30 mg / l titanium dioxide nanoparticles at 24 and 48 hours of exposure time respectively. The highest amount of most polyphenols including rosmarinic acid, cinnamic acid and rutin were achieved during 24 hours of treatment. The highest levels of SO6 protein was obtained in hairy roots treated with 50 mg/l titanium dioxide nanoparticles after 48 hours. By laboratory optimization of hairy root production of soap plant, SO6 can be produced in fermenter.

bacteria species; Burkholderia pseudomallei, Vibrio cholera, and Shigella dysenteriae are the main pathogen causing  Melioidosis, Cholera, and Shigella in humans respectively. bacterial species have a protein that has antigenic prosperities along with being a mucus based adjuvant thus easing their delivery and immunogenicity. These proteins were BLF1(B. pseudomallei), CTxB (V. cholera), and StxB (S. dysenteriae). The goal of this study was a chimeric gene composed of Blf1, CTxB, and StxB and expression of these proteins from E.coli.

Using Bioinfoamtic tools such as Protparam and Modeller the different combinations of the chimeric protein were assessed. The blf1 and stxB genes were PCR amplified from pET28a-BFL1-StxB. The 850bp fragment encompassing BFL1-StxB was cloned using SalI and NotI in the pET28a-CTxB vector. The engineered plasmid was transformed in BL21(DE3). The ctxB-stxB-blf1 gene was expressed following induction with IPTG. This induction was then assessed using SDS-PAGE.

Chimeric protein instability index gets 40.06 and the half-life of it in E. coli was over 10 hours. Codon Adaptive Index (CAI) rise to 0/9 and GC content increase to 54.9. The optimal vaccine combination for the chimeric protein was determined to be CTxB-BLF1-StxB. Expression of recombinant protein in E. coli led to the production of a chimeric protein with 48 kDa molecular weight.

The findings of the current study revealed that this antigen can be raised as a recombinant vaccine candidate which can be due to such factors as a property of the ability to stop protein synthesis in BLF1, CTxB adjuvant, and delivery of StxB protein.