肉类研究Issue(7)：11-15,5.DOI:10.7506/rlyj1001-8123-201507003
不同细胞破碎方法对肉牛、牦牛背最长肌双向电泳图谱的影响
Comparison of Different Cell Disruption Methods for Two-Dimensional Electrophoresis Analysis of Beef and Yak Muscle
摘要
Abstract
Three cell disruptionmethods, i.e., liquid nitrogen grounding, liquid nitrogen grounding aided ultrasonication and liquid nitrogen grounding with glass beads were investigated to ifnd the most appropriate one for the extraction and two-dimensional electrophoresis analysis of proteins inlongissimusdorsi muscles of beef cattle and yak. The same lysis buffer was used in the different cell disruptionmethods and thetwo-dimensional electrophoresis analysis was performed using isoelectricfocusing andsodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) by silver staining. The protein contents oflongissimusdorsi muscles were determined by theBradford method. Analysis of the two-dimensional electrophoresis images was conducted using using PDQuest 8.0.1 software. Results showed that the protein contentsof beef and yaklongissimusdorsi muscles with the three cell disruptionmethods were significantly different, (2.59±0.15) and (2.84 ± 0.29)μg/μL, (2.01 ±0.04) and (2.21 ±0.02)μg/μL, and (1.77± 0.11) and (1.95± 0.19)μg/μL forliquid nitrogen grounding aided ultrasonication, liquid nitrogen grounding with glass beads and liquid nitrogen grounding, respectively. The numbers of protein spots in the electrophoresis images of beeflongissimusdorsiobtained using these three cell disruption methods were (279 ± 19), (581 ±15) and (363 ±21), respectively, whereas those of yaklongissimusdorsi were (292±15), (596 ± 12) and (385 ±17), respectively. Among these three methods,liquid nitrogen grounding aided ultrasonication yielded the highest protein contents, the greatest number of proteins and the best separation of protein spots from beef and yaklongissimusdorsi. No signiifcant differences in the protein contents of beef and yaklongissimusdorsi were observed among the three cell disruption methods.关键词
细胞破碎/肉牛/牦牛/背最长肌/蛋白质提取/双向电泳Key words
cell disruption/beef muscle/yak muscle/longissimus dorsi muscles/protein extraction/two-dimensional electrophoresis分类
轻工纺织引用本文复制引用
左惠心,韩玲,牛克兰,王琳琳,张佳莹,余群力..不同细胞破碎方法对肉牛、牦牛背最长肌双向电泳图谱的影响[J].肉类研究,2015,(7):11-15,5.基金项目
国家自然科学基金地区科学基金项目（31460402）；甘肃省重大科技专项（143NKDP020）；国家现代农业产业（肉牛牦牛）技术体系建设专项 （）
