摘要
物种定量鉴别检测技术,已成为肉类真伪鉴别研究的热点。现有常用的实时荧光PCR法在定量检测方面存在方法缺陷。而基于蛋白质的ELISA检测方法,具有抗干扰能力强、快速、对仪器要求低的优点,已越来越多应用于食品中特征蛋白的定量检测。肉制品加工工艺中,热处理是蛋白质变性的主要原因,因此应选择热稳定特征蛋白,而建立常见肉类热稳定蛋白质双向电泳图谱是选择热稳定特征蛋白的基础。然而双向电泳是一个复杂的过程,想获得良好的电泳图谱并找到特征蛋白点需要多方面进行优化。从蛋白质提取纯化方法、IPG胶条长度、p H范围、上样量选择方面对其进行优化,最终获得稳定的、能够找到特征蛋白点的双向电泳图谱。
Quantitative identification of meat species become an important diagnostic challenge. Normal reatime PCR method has deficiency for quantitative identification. ELISA method based on protein is more suitable for application, because the advantages of strong interference resistance, less time consumption and less requirement of instrument, which is more and more used in food authorities. Heat is the major cause of protein denature during meat processing. Therefore heated-stable protein should be chosen. Construct 2-DE maps of meats is the fundamental step of heated-stable protein chosen. But two-dimensional electrophoresis is complicated, parameters should be optimized. In this study, we optimized protein extract and purified protocol, IPG strip length and pH range, loading amount. Finally, we constructed a solid foundation for the proteomic analysis of meat.
出处
《食品科技》
CAS
北大核心
2016年第5期131-135,共5页
Food Science and Technology
基金
广东省质量技术监督局科技项目(2013CZ05)
