Expression of the Recombinant Protein Containing CfaB, ST, CfaE, and LtB from Enterotoxigenic Escherichia coli and Loading It in Chitosan Nanoparticles | ||
| Pathobiology Reserach | ||
| Article 2, Volume 22, Issue 2, 2019, Pages 69-75 PDF (647.78 K) | ||
| Document Type: Original Research | ||
| Authors | ||
| Z.S. Hosseini1; J. Amani* 2; F. Hosseini1 | ||
| 1Cell & Molecular Biology-Biotechnology Department, Biological Sciences Faculty, Tehran North Branch, Islamic Azad University, Tehran, Iran | ||
| 2Applied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran | ||
| Abstract | ||
| Aims: Enterotoxigenic Escherichia coli (ETEC) is the most important bacteria causing traveler’s diarrhea. The bacterium has several virulence factors, including colonization factors (CFs) or Escherichia coli adhesins, heat-labile (LT), and heat-stable (ST) toxins. The design and production of vaccine against this disease is one of the goals of the World Health Organization due to increased antibiotic resistance and a reduction of healthy water sources. An effective subunit vaccine against ETEC could include a toxoid from both toxins and colonization factors. The aim of the current study was to express, purify, and encapsulate the recombinant protein in chitosan nanoparticles. Materials and Methods: In the present experimental study, the E. coli BL21DE3 harbring pET-28a-cscl vector was used. The chimeric cscl gene is composed of cfab along with st toxin, cfae, and ltb. After the expression and purification of recombinant protein, using Ni-NTA column, Western blotting was performed with anti-His antibody. Then, the CSCl protein was encapsulated in chitosan nanoparticles and the particle size was measured. Findings: The recombinant CSCL protein was purified by Ni-NTA column and urea denaturation method. Then, this purified protein (~57kDa) was confirmed by Western blotting and the size of the nanoparticles was estimated as 112.0 nm with 98.8% of encapsulation efficiency. Conclusion: With some advantages, including the presence of surface and important antigens of ETEC and encapsulating in chitosan nanoparticles, the CSCL recombinant protein can be considered as a candidate for producing oral nanovaccine and stimulating of mucosal and systemic immune response. | ||
| Keywords | ||
| Enterotoxigenic Escherichia coli; Heat-labile toxin; Heat-stable toxin; Chitosan nanoparticles; Escherichia coli adhesins; Traveler’s diarrhea | ||
| References | ||
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