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KMID : 0380420200440030104
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Journal of Prventive Veterinary Medicine
2020 Volume.44 No. 3 p.104 ~ p.112
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Construction of Salmonella Enteritidis ghosts manipulated by the λ phage holin-endolysin system that express HA1 domain of hemagglutinin of influenza H1N1 virus
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Won Ga-Yeon
Lee John-Hwa
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Abstract
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Salmonella Enteritidis (SE) infection, one of a major foodborne disease remains considerable public health concerns globally. In this study, we have constructed that the genetically inactivated SE ghost vaccine candidate, JOL2220 (Δlon ΔcpxR Δasd), which express the immunogenic hemagglutinin portion (HA1) of H1N1 virus on the Salmonella surface. The lysis gene cassette, holin-endolysin system originated from double DNA bacteriophage λ was cloned into the ghost vector plasmid pJHL464, which activated under the control of the convergent promoter system. Scanning electronic microscopic examination revealed that transmembrane tunnels of JOL2220 was formed following the 48 hr of the lysis mediated by the holin-endolysin system. In vitro expression of HA1 antigenic protein was also determined by Western blot analysis. The Chickens immunized with JOL2220 ghost via intramuscular and oral routes showed significant increase in IgG level against both SE and HA1 antigen. Further, we observed markedly enhancement of lymphocyte proliferation and T-cell differentiation in the primed PBMC of the immunized chicken, compared to those in the control group. Collectively, the results indicated that the SE ghosts expressing HA1 antigen may have immunostimulatory properties against Salmonella and influenza infection where the chicken is the predominant reservoir.
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KEYWORD
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Salmonella Enteritidis, foodborne disease, holin-endolysis system, the bacterial ghost, HA1 protein
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