| Analysis of outer mombrane proteins of Brucella abortus using two dimensional polyacrylamide gel electrophoresis |
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Byung-su Kim1, Sun-hee Kim1, Jong-suk Kim2, Byeong-kirl Baek3 |
1Department of Clinical Pathology, Kunsan Junior College
2Department of Biochemistry, Medical School Chonbuk National University
3College of Veterinary Medicine, Chonbuk National University |
| 2차원 전기영동법을 이용한 Brucella abortus 세포외막 특이단백질의 분석 |
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김병수1, 김선희1, 김종석2, 백병걸3 |
1군산전문대 임상병리과
2전북의대생화학교실
3전북대학교 수의학과 |
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| Abstract |
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Outer membrane proteins(OMPs) of Brucella abortus 1119-3 strain were extracted by Triton X-100 treatment, and fractionated by DEAE-cellulose column chromatography and Sephacryl S-300 column chromatography. The antigenic proteins in these fractions were identified by Western blot analysis. In Western blot analysis, a single band(38kDa) was observed in the DEAE fractions from 36th fraction to 38th fraction against sera of cattle infected with B abortus. And other fractions have several bands. However, the Sephacryl S-300 fractions exhibited a total of 3 peaks of proteins with a broad range from about 30 to 116kDa. In order to characterize further, the extracted OMPs and the DEAE fractions were analyzed by two dimensional polyacrylamide gel electrophoresis(2-DE) and Western blot using serum from naturally infected cattle with Brucella spp. The 2-DE immunoblots of DEAE fraction showed immunoreactive spots more than twenty two. The major protein spots have ranging from about 32 to 47kDa. The pI values of the spots were detected from pH 4.7 to 5.4. Among the major protein spots, the 38kDa protein which is a specific antigen, located at the point of approximately a pI 4.8. |
| Key Words:
Brucella abortus, OMPs, colwnn chromatography, two dimensional electrophoresis |
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