전북지역 한우에서 분리한 Theileria sergenti의 항원성에 관한 연구

다국어 초록 (Multilingual Abstract)

A splenectomized 5 monthold calf was inoculated with cryopreserved Theileria sergenti infected korean native cattle in chonbuk district.
At peak parasitemia(40%), blood was collected, washed and then theileria sergenti was obserbed to delineate the morphologic characteristic with transmission electron microscope.
Also, T.sergenti merozoite was lysed with infected RBC and then isolated by differential centrifugation. Antigenic profile of isolated T.sergenti organism was analyzed by SDS-PAGE and western blotting technique.
The result could be summarized as follows:
1. light microscopic observation
Micrgraph of the infected erythrocyte with Theileria Sergenti with Giemsa stain(×1000.)
T.sergenti of commet form(48%), oval form(20%), dot form(12%), baccilus form(11%), and the other form(9%) are observed on infected erythrocytes.
2. trasmission electron microscopic observation:
T.sergenti merozoite were observed on numerous shapes, also cytoplasm cotained rhoptris, food vacuole, and tubule.
Merozoite varied from 1.7-2um in diameter, divided by a binary fission. The most infected erythrocytes cotained veil (hemosiderin)
3. SDS-polyacrylamide gel electrophoresis
T.sergenti merozoit was subjected to electrophoresis in a 12% polyacrylamide gel. The gel was stained with 0.1% Coomassian brilliant blue R to visualize protein band. Stained gel revealed at least twelve protein bands of approximately 14Kd, 28Kd, 30Kd, 34Kd, 36Kd, 41Kd, 56Kd, 66Kd, 72kd, 97Kd and 116Kd but normal RBC membrane revealed protein bands of 30-34Kd, 36Kd, 42Kd, 58Kd, 66Kd, 97Kd and 116Kd.
4. Enzyme linked immunoelectrotransfer blot(western blot)
T.sergenti antigen recognized by specific anti- T.sergenti antibodies demonstrated 28Kd, 30Kd, 38Kd, 56Kd, 58Kd, 66Kd, 97Kd and 116Kd molecular weight range.
Negative reactivities were observed in normal bovine serum with T.sergenti and normal erythrocytic antigens. Predominant protein of T.sergenti merozoite antigen were founded at 28Kd, 30Kd and 41Kd molecular weight. On going studies will analyze the relatively inprtance of those antigens for immunization of T.sergenti in Korean native cattle.

목차 (Table of Contents)

• 목차
• 영문초록 = ii
• 서론 = 1
• 재료 및 방법
• 1. Theileria sergenti의 증균 = 3
• 2. 전자현미경 관찰 = 3
• 3. 항원준비 = 4
• 4. T.sergenti에 대한 양성 및 음성혈청 준비 = 4
• 5. SDS-PAGE = 5
• 6. western blot = 6
• 결과
• 1. T.sergenti원충의 광학 및 전자현미경적 관찰 = 7
• 2. T.sergenti항원의 SDS-PAGE = 8
• 3. western blot 소견 = 9
• 고찰 = 10
• 결론 = 15
• 참고문헌 = 16
• 사진설명 = 21
• 사진부도 = 23