农业生物技术学报2012，Vol.20Issue(1)：105-112,8.DOI:10.3969/j.issn.1674-7968.2012.01.014
猪肌肉组织双向电泳分离条件的建立及常见问题分析
Establishment and Problem Analysis of Two-dimensional Electrophoresis Separation on Porcine Skeletal Muscle
摘要
Abstract
To establish two-dimensional gel electrophoresis methods (2-DE) for porcine skeletal muscle proteome, the experimental conditions of 2-DE was optimized, and various extraction methods, different loading quantities and pH IPG strips, different adhesive concentration and gel staining were studied. The results showed that the process of grinding in liquid nitrogen and following by ultrasonication was more appropriate than that of grinding in liquid nitrogen, which could obtained better 2-DE map for low nucleic acid contamination and more protein spots. Compared with different pH IPG strips, 18 cm pH 4-7 IPG strips were better than pH 3-10 NL. For silver staining of 18 cm pH 4-7 IPG strip, the suitable loading quantity was 150 |xg. Furthermore, the reproducibility of three gels in same sample was more 70%. Meanwhile, the common problems occurring in 2-DE experiments were analyzed. The results indicated that the 2-DE maps of porcine skeletal muscle were obtained with high resolution and reproducibility, and can be used for the further proteome analysis.关键词
猪/肌肉/双向电泳/蛋白质组学/分析Key words
Pig/Skeletal muscle/Two-dimensional electrophoresis/Proteomics/Analysis引用本文复制引用
徐永杰,熊远著..猪肌肉组织双向电泳分离条件的建立及常见问题分析[J].农业生物技术学报,2012,20(1):105-112,8.基金项目
国家重点基础研究发展规划(973)项目(No.2006CB 102102)、转基因生物新品种培育重大专项(No.2008ZX08006-002)和信阳师范学院青年骨干教师资助计划(2011) （973）
