1.Effective methods for the production of immunoglobulin Y using immunogens of Bordetella bronchiseptica, Pasteurella multocida and Actinobacillus pleuropneumoniae.
Na Ri SHIN ; In Soo CHOI ; Jong Man KIM ; Won HUR ; Han Sang YOO
Journal of Veterinary Science 2002;3(1):47-57
Swine respiratory diseases induce severe economic losses in the swine industry worldwide. Several methods have been developed and applied to control these diseases. However, there are still problems of disease control in the swine industry. Recently, egg yolk antibodies have been found to offer several advantages for disease control in animals and humans. In a previous study (24), antibodies to several causative pathogens of swine respiratory diseases were developed. However, several problems remained, especially in terms of reduced laying rates. Therefore, experimental vaccines were reformulated with various bacterial antigens of the swine respiratory diseases. After immunizing hens with the antigens, antibody profiles and other effects including laying rates were investigated and compared to those of the previous study. Profiles of antibody titers were very similar with those of the previous study. However, side effects, such as depression, weakness, reduction of laying rates and mortality, were dramatically lowered and laying rates were increased in hens injected with certain experimental vaccines. In particular, laying rates of hens injected with vaccines against atrophic rhinitis were increased up to 84% by injecting a vaccine composed of only the DNTs of B. bronchiseptica and P. multocida D:4. Efficacies of the vaccines against swine pneumonic pasteurellosis and pleuropneumonia were very similar with those of the previous study. These results suggest that new vaccines could be effective in the production of egg yolk antibodies against the causative agents of swine respiratory diseases.
Actinobacillus pleuropneumoniae/classification/genetics/*immunology
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Animals
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Antibodies, Bacterial
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Antibody Formation
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Bacterial Outer Membrane Proteins/genetics/isolation & purification
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Bordetella bronchiseptica/classification/genetics/*immunology
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Egg Yolk/microbiology
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Female
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Immunoglobulins/*genetics
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Oviposition
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Pasteurella multocida/classification/genetics/*immunology
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Serotyping
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Swine
2.Expression of apxIA of Actinobacillus pleuropneumoniae in Saccharomyces cerevisiae.
Sung Jae SHIN ; Jong Lye BAE ; Young Wook CHO ; Moon Sik YANG ; Dae Hyuk KIM ; Yong Suk JANG ; Han Sang YOO
Journal of Veterinary Science 2003;4(3):225-228
Actinobacillus pleuropneumoniae is an important primary pathogen in pigs, in which it causes a highly contagious pleuropneumoniae. In our previous study, apxIA gene amplified from A. pleuropneumoniae Korean isolate by PCR with primer designed based on the N- and C-terminal of the toxin was cloned in TA cloning vector and sequenced. The nucleotide sequences of apxIA gene was reported to GeneBank with the accession numbers of AF363361. Identity of the Apx IA from the cloned gene in E. coli was proved by SDS-PAGE and Western blot. Yeast has been demonstrated to be an excellent host for the expression of recombinant proteins with uses in diagnostics, therapeutics and vaccine productions. Therefore, to use the yeast as a delivery system in new oral subunit vaccine, apxIA gene was subcloned into Saccharomyces cerevisiae, and ientified the expression of Apx IA protein. First, apxIA gene was amplified by PCR with the primers containing BamHI and SalI site at each end. Second, the DNA digested with BamHI and SalI was ligated into YEpGPD-TER vector, and transformed into S. cerevisiae 2805. Third, after identification of the correctly oriented clone, the 120-kDa of Apx IA protein expressed in S. cerevisiae 2805 was identified by SDS-PAGE and Western blot.
Actinobacillus pleuropneumoniae/*genetics/isolation & purification/metabolism
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Animals
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Bacterial Proteins/*biosynthesis/genetics
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Blotting, Western/veterinary
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Cloning, Molecular
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DNA, Bacterial/chemistry/genetics
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Electrophoresis, Polyacrylamide Gel/veterinary
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Hemolysin Proteins
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Pleuropneumonia, Contagious/microbiology
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Polymerase Chain Reaction/veterinary
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Saccharomyces cerevisiae/genetics/*metabolism
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Swine
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Swine Diseases/microbiology