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자료유형
학술저널
저자정보
정지아 (농림축산검역본부) 김하영 (농림축산검역본부) 김종완 (농림축산검역본부) 이완규 (충북대학교) 변재원 (농림축산검역본부)
저널정보
충북대학교 동물의학연구소 Journal of Biomedical and Translational Research Journal of Biomedical and Translational Research 제18권 제3호
발행연도
2017.9
수록면
103 - 107 (5page)

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It has been known that the ability of Shiga toxin-producing Escherichia coli (STEC) to produce Stx2e in culture media plays a role in the diagnosis of edema disease and determination of subunit vaccine candidates in STEC isolates. To examine the efficiency of Stx2e production in several commercial media, a Stx2e-producing strain (KEFS1302) was grown in four different media: ISO-Sensitest broth (ISB), E. coli broth (ECB), trypticase soy broth (TSB), and Mueller Hinton broth (MHB), with or without mitomycin C at 37°C (250 rpm) for 6 h. Toxin production was measured by enzyme-linked immunosorbent assay. In the presence of mitomycin C, ECB was found to be the most suitable medium, reaching a production peak (OD600 = 1.2) at 1 h; Stx2e was mostly produced during the logarithmic phase (within 3 h). On the other hand, toxin production in ISB reached a peak at 3 h after incubation in the absence of mitomycin C. Stx2e was purified by fast protein liquid chromatography (FPLC) using anion-exchange chromatography. The 43 kDa band of Stx2e was confirmed by western blot using the ECB supernatant. Our results showed that ECB and ISB media would be a suitable medium for mass production of Stx2e even if the toxin production is dependent on time.

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