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KMID : 1007520100190041107
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Food Science and Biotechnology
2010 Volume.19 No. 4 p.1107 ~ p.1112
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Buffer Optimization for Bovine Longissimus Muscle Tissues: Proteome Analysis of Korean Native Cattle Using 2-Dimensional Gel Electrophoresis
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Lee Hong-gu
Han Jeung-a
Lee Ki-Bum
Kim Eun-bea
Jin Yong-Cheng
Oh Jin-Ju
Hwang Jin-hee
Kang Han-suk
Kim Sang-hun
Seo Kang-suk
Kang Sang-Kee
Choi Yun-Jaie
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Abstract
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Because beef contains a high proportion of marbling, we sought to optimize the solubilization of the longissimus muscle proteome using modified lysis and isoelectric focusing (IEF) rehydration buffers. Of the 3 lysis buffers tested, those containing a thiourea-urea mixture provided superior resolution, whereas that which contained only urea yielded consistently poor results. In addition, we found that the IEF rehydration buffer containing a thiourea-urea mixture, a high dithiothreitol level, and Pharmalyte was able to generate sharper 2- dimensional (2-D) gels, whereas that which contained immobilized pH gradient (IPG) buffer had no an effect on protein resolution. The number of different protein spots between the muscle-development stage (11 months) and the fat-development stage (17 months) in Hanwoo steers was 39, whereas the use of normal buffer resulted in a detection of a difference of only 7 protein spots. This study provides a methodological tool for studying the proteome of bovine longissimus muscle with a high fat content using 2-D gel electrophoresis (2-DE) gels.
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KEYWORD
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bovine longissimus muscle, 2-dimensional gel electrophoresis (2-DE), 2-DE analysis, solubilization, resolution
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