• 제목/요약/키워드: turbid plate

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Tyrosine Phenol-Lyase의 고속탐색기술 개발 및 방향성 분자진화 (High Throughput Screening and Directed Evolution of Tyrosine Phenol-Lyase)

  • 최수림;나유진;김도영;송재준;홍승표;성문희;이승구
    • 한국미생물·생명공학회지
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    • 제34권1호
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    • pp.58-62
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    • 2006
  • 티로신 및 방향족 아미노산 유도체의 생물학적 합성에 이용되는 유용 효소인 tyrosine phenol-lyase(TPL) 활성을 평판배지에서 정량적으로 감지할 수 있는 기술을 개발하였다. 불투명 평판배지(turbid plate)의 제조과정은 먼저, 2 N염산과 DMSO로 조성된 공용매에 난용성 티로신을 400 mM 농도로 녹여서 배지에 가하고, 온도를 조절하여 니들형 티로신을 형성하도록 결정화를 유도하는 것이었다. 니들형 티로신은 일반적인 컬럼형에 비하여 약 $5{\sim}6$배 낮은 농도인 3.6g/L에서도 불투명 평판배지를 제조할 수 있었으며, 효소활성에 의하여 쉽게 분해되고, TPL활성의 고감도 고속탐색기술 개발에 적합하였다. 이 불투명 평판배지에 변이유발PCR법으로 제조한 TPL 라이브러리를 도말하고, 단일 콜로니 주변에서 형성되는 투명환의 크기와 실측한 TPL활성을 비교한 결과 직접적인 비례관계가 있음을 확인하였다. 따라서 난용성 물질의 미세입자를 평판배지에서 직접 발생시킨 후, 효소활성에 의한 투명환의 형성을 정량 관찰하는 본 연구의 방법은 신규 고활성 TPL의 분리 및 방향성 분자진화 등을 위한 고속탐색기술에 유용하게 사용될 수 있다.

Fatal Peritonitis associated with Pasteurella multocida in an Asian Small-Clawed Otter (Aonyx cinereus)

  • Kyung-Seok Na;Hyoung-Seok Yang;Won-Hee Hong;Jae-Hoon Kim
    • 한국임상수의학회지
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    • 제41권1호
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    • pp.54-59
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    • 2024
  • A 12-year-old female small-clawed Asian otter (Anoyx cinereus) with a one-week history of anorexia, chills, and abdominal distension was found dead. Grossly, yellowish-brown turbid fluids accumulated in abdominal cavity of the otter, and yellowish thread-like fibrinous materials were found on the surface of abdominal organs. Several variable sized yellowish-white crystalloids were scattered on the medullary space of kidneys. Histologically, diffuse serositis (peritonitis) characterized by the fibrinous exudates, thickened serosal capsule and the swelling of mesothelial cells were observed in the serosa of liver, spleen, stomach, and intestine. Multifocal necrosis, hemorrhage, infiltration of macrophage, and brown pigments were presented in the liver. Isolated bacteria from ascites and fibrinous materials in abdominal visceral surface were white, smooth and convex with characteristic mousy odor on blood agar plate. These bacteria were confirmed as Pasteurella (P.) multocida type A by polymerase chain reaction analysis. Based on the gross examination, histopathologic findings and bacterial experiments, this otter was diagnosed as severe peritonitis associated with P. multocida and necrotic hepatitis.

Lactobacillus casei Bacperiophage의 분류 및 특성에 관한 연구 (Classification and Characterization of Bacteriophages of Lectobacillus casei)

  • 김영창;박민철;강국희;윤영호;이광웅
    • 미생물학회지
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    • 제17권4호
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    • pp.165-178
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    • 1979
  • Phages of Lactobaciilus casei (PLC) isolated from plant drainage were classified and characterized. The results are as follows : 1. On the basis of host range pattern, phages could be divided into 2 groups (PLC-B and PLC-C). PLC-B group phages could be further divided into 5 sub-groups $(B_1, \;B_2, \;B_3, \;B_4, \;and\;B_5)$. Although PLC-C group phages had the same host range, they could be also divided into 2 sub-groups $(C_1\;and\;C_2)$ by morphlogical type. 2. It was $B_3$ group phages that represented a major proportion (44.4%) of phages tested. However, $B_1$ group phages were shown to have the widest host range. 3. Electron micrographs revealed that the phages fell into three different morphological types. $(B_1, \;B_2, \;and\;B_3)$ group phages hd a hexagonal head (52nm in diameter) and a sheathless noncontractile (245 nm in length). $B_4\;and\;C_2$ group phages had a hexagonal head (56 nm) and a short flexible tail (169nm) having no sheath. $B_5\;and\;C_1$ group phages were shown to have a hexagonal head (81 nm) and a contractile tail (140 nm) having a sheath, a base plate and tail fibers. 4. The inactivation of the phages by antisera indicated that serological relationships correlated completely with morphological types. 5. $B_1, \;C_1\;and\;C_2$ group phages produced a large (1, 2 mm in diameter) plaque with a clear ring. The morphology of plaques of $B_3\;and\;B_5$ group phages was the same as those produced by the above, but the average plaque sizes for $B_3\;and\;B_5$ were 0.8 mm abd 0.5 mm, respectively. $B_2\;and\;B_4$ group phages produced a small (0.5 mm) turbid plaque with an irregular edge. 6. The latent period and the average burst size of $B_1\;and\;B_3$ group phages were 90 min and 100, respectively. These phages reuqired calcium ions for their miltiplication. 7. $B_3$ group phages could not be absrobed to R-variant $KC_1$. 8. The order of resistance of phages to heat was $B_2\;>\;B_1, B_4\;and\;B_5\;>\;B_3\;and\;C_2, \;B_5$ group phages were more stable than $B_3$ in various pH values. $C_2$ group phages were more sensitive to UV irradiation than $B_1\;and\;B_3$ group phages. 9. Strains YIT9018 and IAM 1043 were induced by mitomycin C treatment. Phage particles detected in the lysates had a hexagonal head (38 and 49 nm, respectively), but no tail. Any sensitive indicator strain could not be isolated in spite of repaeated trials.

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