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Expression of Recombinant Porcine Interleukin-2 and Application ofIts Antibody to Immunoassays
유한상,In-Soo Chol 대한수의학회 2002 Journal of Veterinary Science Vol.3 No.3
Interleukin-2 plays an important role in Tlymphocyte proliferation and imune response regul-ations. In this tudy, porcine IL-2 cDNA was clonedfrom peripheral blood mononuclear cells, and recom-binant porcine IL-2 (rpIL-2) was expresed in Escherichiacoli . The size of rpIL-2 without signal peptides wasabout 15 kDa when determined by SDS-PAGE andproduced from ice imunized with the purifiedrpIL-2, and its pecificity was examined by Westernblotting and ELISA. In the Western blotting asay,anti-rpIL-2 and anti-recombinant human IL-2 (rhIL-2)antibodies pecifically recognized rpIL-2 and rhIL-2,respectively. However, anti-rpIL-2 antibody did notrecognize rhIL-2, and anti-rhIL-2 antibody also didnot react with rpIL-2 in the same asay. In ELISA,anti-rpIL-2 antibody strongly interacted with bothrpIL-2 and rhIL-2, and anti-rhIL-2 antibody also efici-specificity of anti-rpIL-2 antibody for pIL-2 wasdemonstrated by Western blotting and ELISA. It wasalso shown that ELISA is more efficient han Westernblotting in determing the species cros-reactivity ofanti-rpIL-2 antibody.
유한상,Deog Yong Lee,Young Wook Cho,강상균,신성재 대한수의학회 2004 Journal of Veterinary Science Vol.5 No.4
Interleukin-6 (IL-6) is introduced as a marker of disease. At present, a variety of method may be used to quantify expression of this protein. Antigen capture-ELISA is a sensitive and accurate quantification method previously used with ovine, rat, and human IL-6 proteins. However, it has never been reported to quantify porcine IL-6 protein using capture ELISA. In this study, we generated and characterized a set of IgY and monospecific polyclonal antibodies to recombinant porcine IL-6 (rpIL-6), and combining these with a sensitive and specific capture-ELISA for a diagnostic purpose. cDNA encoding the mature protein coding region of porcine IL-6 was cloned and expressed with pQE-30UA expression vector. rpIL-6 was then expressed and purified by using Ni-NTA resin. Protein mass of 24 kDa was found with SDS-PAGE and the identity of the protein was confirmed by Westernblot. Production of polyclonal antibodies against rpIL-6 was performed using the purified rpIL-6 in mice and hens. An antigen capture-ELISA was developed with the antibodies after their extraction. To compare the IL-6 level in the different sanitary state of farms, pig sera were randomly collected and concentration of IL-6 in the sera was measured with the antigen capture-ELISA. The capture-ELISA with the optimal concentration of antibodies, in this study, was able to detect about 10 ng/ml of rpIL-6. IL-6 levels determined with the capture-ELISA in pig sera showed positive correlation with the sanitary states of the farms. These results suggested that the developed antigen capture-ELISA could be a good tool for the screening of microbial infection in pig farms.
COVID-19 and Veterinarians for One Health, Zoonotic- and Reverse-Zoonotic Transmissions
유한상,Dongwan Yoo 대한수의학회 2020 Journal of Veterinary Science Vol.21 No.3
A novel coronavirus emerged in human populations and spread rapidly to cause the global coronavirus disease 2019 pandemic. Although the origin of the associated virus (severe acute respiratory syndrome coronavirus 2 [SARS-CoV-2]) remains unclear, genetic evidence suggests that bats are a reservoir host of the virus, and pangolins are a probable intermediate. SARS-CoV-2 has crossed the species barrier to infect humans and other animal species, and infected humans can facilitate reverse-zoonotic transmission to animals. Considering the rapidly changing interconnections among people, animals, and ecosystems, traditional roles of veterinarians should evolve to include transdisciplinary roles.
유한상,강상균,Deog-Yong Lee,Sung-Jae Shin,Jeong-Min Ahn 대한수의학회 2005 Journal of Veterinary Science Vol.6 No.3
The worldwide use of antimicrobials in different fields has created enormous pressure for the selection of resistance among opportunistic bacterial pathogen. One hundred four E. coli isolates were collected and identified from swine with diarrhea in Korea during the period of 2002. The isolates showed highly resistant to streptomycin (99.0%), tetracycline (97.1%), neomycin (91.3%) and carbenicillin (84.6%) in antimicrobial susceptibility test. Moreover, all of the isolates showed multiple antimicrobial resistant to more than 3, and 85% of them were resistant to more than 7 of total 14 antimicrobial agents. In comparison with isolates in 1998, resistance to antimicrobials was more frequent among the isolates in 2002. Presence of class 1 integrons was investigated through amplification of the gene with PCR, and could be classified 8 groups by pattern of 4 different amplicons. Class 1 integrons were observed in 67 strains (64.2%) of E. coli from swine in Korea. One and 1.6 kbp of amplicons were revealed to contain aadA1 and aadB-aadA1 gene cassettes respectively. Two kbp of amplicon had three different gene cassettes, dhfrXII-orfF-aadA2, and 3.0 kbp of amplicon includes aadB-cmlA1 gene cassettes. The worldwide use of antimicrobials in different fields has created enormous pressure for the selection of resistance among opportunistic bacterial pathogen. One hundred four E. coli isolates were collected and identified from swine with diarrhea in Korea during the period of 2002. The isolates showed highly resistant to streptomycin (99.0%), tetracycline (97.1%), neomycin (91.3%) and carbenicillin (84.6%) in antimicrobial susceptibility test. Moreover, all of the isolates showed multiple antimicrobial resistant to more than 3, and 85% of them were resistant to more than 7 of total 14 antimicrobial agents. In comparison with isolates in 1998, resistance to antimicrobials was more frequent among the isolates in 2002. Presence of class 1 integrons was investigated through amplification of the gene with PCR, and could be classified 8 groups by pattern of 4 different amplicons. Class 1 integrons were observed in 67 strains (64.2%) of E. coli from swine in Korea. One and 1.6 kbp of amplicons were revealed to contain aadA1 and aadB-aadA1 gene cassettes respectively. Two kbp of amplicon had three different gene cassettes, dhfrXII-orfF-aadA2, and 3.0 kbp of amplicon includes aadB-cmlA1 gene cassettes.