TY - JOUR ID - 5259 TI - Evaluation of immune response to recombinant Bacillus anthracis LFD1-PA4 chimeric protein JO - Iranian Journal of Veterinary Research JA - IJVR LA - en SN - 1728-1997 AU - Mirhaj, H. AU - Honari, H. AU - Zamani, E. AD - Ph.D. Student in Nano Biotechnology, Department of Biology, Faculty of Basic Science, Imam Hossein University, Tehran, Iran AD - Department of Biology, Faculty of Basic Science, Imam Hossein University, Tehran, Iran AD - MSc Student in Cellular and Molecular Biology, Department of Biology, Faculty of Basic Science, Imam Hossein University, Tehran, Iran Y1 - 2019 PY - 2019 VL - 20 IS - 2 SP - 112 EP - 119 KW - Anthrax KW - Antibody titer KW - LFD1 KW - LFD1-PA4 KW - PA4 DO - 10.22099/ijvr.2019.5259 N2 - Background: Anthrax is a particularly dangerous infectious disease that affects humans and livestock. Efficacious vaccines that can rapidly induce a long-term immune response are required to prevent anthrax infection in humans. Domains 4 and 1 of the protective antigen (PA) and lethal factor (LF), respectively, have very high antigenic properties. Aims: In this experimental study, the pET28a-lfD1-pa4 expression vector was designed, constructed and transferred into E. coli BL21 (DE3) plysS. Methods: For this purpose, pa4 gene was amplified by polymerase chain reaction (PCR) and cloned in a pGEM T-easy vector. The pGEM-pa4 and pGEM-lfD1 were digested by XbaI and HindIII enzymes. The ligation reaction was performed by ligase T4 enzyme and the gene cassette, lfD1-pa4, was subcloned in pET28a and transferred to E. coli BL21 (DE3) PlysS. Expression and purification of chimeric proteins were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting techniques. The chimera LFD1-PA4 and mixed LFD1+PA4 proteins were injected four times into mice and antibody production was relativity evaluated by enzyme-linked immunosorbent assay (ELISA) test. Results: The results showed that both chimeric and mixed proteins are immunogenic, but LFD1-PA4 has a higher potential to stimulate mice immune system. Conclusion: LFD1-PA4 chimeric protein induced a higher immune response than LFD1+PA4 mixed protein and elicited antibody responses to LF and edema factor (EF), therefore, it holds promise to be a more effective trivalent vaccine candidate to use in anthrax prevention. UR - https://ijvr.shirazu.ac.ir/article_5259.html L1 - https://ijvr.shirazu.ac.ir/article_5259_e46b0f4fb04b3bbfe6ec2e38903c65f5.pdf ER -