Buffer Optimization for Bovine Longissimus Muscle Tissues: Proteome Analysis of Korean Native Cattle Using 2-Dimensional Gel Electrophoresis

被引:3
|
作者
Lee, Honggu [2 ,6 ]
Han, Jeunga [1 ]
Lee, Kibum [3 ]
Kim, Eunbea [1 ]
Jin, Yongcheng [1 ,2 ,6 ]
Oh, Jinju [2 ,6 ]
Hwang, Jinhee [2 ,6 ]
Kang, Hansuk [2 ,6 ]
Kim, Sanghun [4 ]
Seo, Kangsuk [5 ]
Kang, Sangkee [1 ]
Choi, Yunjaie [1 ]
机构
[1] Seoul Natl Univ, Dept Anim Sci, Sch Agr Biotechnol, Seoul 151742, South Korea
[2] Pusan Natl Univ, Dept Anim Sci, Miryang 627706, Gyeongnam, South Korea
[3] SongDo Techno Pk, Dept Biotechnol, Inchon 406840, South Korea
[4] Kyung Hee Univ, Dept Biol, Seoul 130701, South Korea
[5] Suncheon Natl Univ, Dept Breeding, Sunchon 540742, Jeonnam, South Korea
[6] Pusan Natl Univ, PNU Special Anim Biotechnol Ctr, Miryang 627706, Gyeongnam, South Korea
关键词
bovine longissimus muscle; 2-dimensional gel electrophoresis (2-DE); 2-DE analysis; solubilization; resolution; CAPILLARY-ZONE-ELECTROPHORESIS; IMMOBILIZED PH GRADIENTS; BACKGROUND ELECTROLYTES; MEMBRANE-PROTEINS; SOLUBILIZATION; PLASMA; MAP;
D O I
10.1007/s10068-010-0157-0
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
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.
引用
收藏
页码:1107 / 1112
页数:6
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