乳及乳制品中乳铁蛋白含量的电泳迁移率检测  

Determination of Bovine Lactoferrin in Milk and Dairy Products by Electrophoretic Mobility Shift Assay

作　　者：黄锐 孙娜娜 张涵 杨孟迪 刘金虎 徐秦峰 HUANG Rui;SUN Nana;ZHANG Han;YANG Mengdi;LIU Jinhu;XU Qinfeng(Zhongken Huashan Mu Dairy Co,Ltd.,Weinan 714000,China;School of Food and Biological Engineering,National R&D Center for Goat Dairy Products Processing Technology,Shaanxi University of Science&Technology,Xi'an 710021,China)

机构地区：[1]中垦华山牧乳业有限公司,陕西渭南714000 [2]陕西科技大学食品与生物工程学院国家羊乳制品加工技术研发专业中心,西安710021

出　　处：《中国乳品工业》2021年第1期46-50,共5页China Dairy Industry

基　　金：陕西省科技厅项目(2018KJXX-043,2019NY-126)。

摘　　要：凝胶电泳迁移率(EMSA)是研究蛋白质和DNA分子相互作用的经典分子生物学方法。本研究根据乳铁蛋白(Lactoferrin,Lf)和DNA分子的特异性结合,基于Lf-DNA复合物和DNA分子迁移率差异,建立了一种聚丙烯酰胺凝胶电泳(PAGE)检测乳铁蛋白的方法。实验中以FAM荧光基团标记DNA分子,根据凝胶成像图中Lf-DNA复合物荧光条带的存在与否、荧光条带灰度值的大小,来判断样品中乳铁蛋白是否存在、含量多少,从而实现乳铁蛋白的定性与定量检测。结果表明,该方法具有良好的特异性及准确性,经过简单稀释后便可直接应用于乳及乳制品中乳铁蛋白的准确定量检测。Electrophoretic mobility shift assay(EMSA)is a classical molecular biological method for studying protein-DNA interactions.Based on the specific binding of bovine lactoferrin(Lf)and DNA molecules,a polyacrylamide gel electrophoresis(PAGE)method for the detection of lactoferrin was developed by using the different mobility between Lf-DNA complex and free DNA.In the experiment,specific DNA was labeled with FAM fluorescent reporter,and the presence of lactoferrin and the concentration of lactoferrin in the sample can be determined according to the existence of Lf-DNA complex band in the gel image and the fluorescence intensity of complex band,respectively.The results showed that this method has good specificity and accuracy for the qualitative and quantitative detection of lactoferrin.Only simple dilution of sample was required for the accuratedetermination of lactoferrin in milk and dairy products.

关 键 词：乳铁蛋白检测 PAGE 电泳迁移率分析 DNA-蛋白质相互作用 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]