摘要
蛋白酶K常用于核酸提取实验中,其作用为去除基因组中的组蛋白以及去除核酸中的DNA酶和RNA酶。目前已有许多国家标准采用分子生物学技术检测食品中的转基因成分、动物属性、食源性致病菌、病毒等,其中核酸提取均需使用蛋白酶K。然而由于目前缺乏统一的蛋白酶K活性检测标准,各个厂家均有各自的活性检测方法,其采用的底物、缓冲液、pH值、温度均大不相同,这也导致了酶活性定义大不相同,这使得所购买的蛋白酶K酶活性不统一,间接也影响了核酸提取的质量导致影响了检测结果。在现有国标蛋白酶活性检测标准基础上对检测方法进行改进,使其更接近使用的条件和温度,为标准建立提供参考。
Proteinase K is commonly used in nucleic acid extraction experiment.Its role is to remove the histone from genome and inactivate Danas and RNase.There are many national standards in china using molecular techniques for the detection of genetically modified organism,animal-originated detection,foodborne pathogens,and virus in food.Which the nucleic acid extraction is required the use of protease K.However,due to the current lack of unified protease K activity detection standards,and manufacturers have their own activity detection method,which adopts various substrate,buffer solution,pH value and temperature,resulted in enzyme activity in different definition.This led to the activity of protease K purchased is not uniform,which effects nucleic acid extraction quality and affects the detection results.In this study we improved the national standard protease activity detecting method,which is more close to the conditions of use,in order to provide a reference as the standard was established.
出处
《食品科技》
CAS
北大核心
2013年第2期266-271,共6页
Food Science and Technology
基金
"十一五"国家科技支撑计划项目(2009BAK61B04)
关键词
蛋白酶K
活性检测
丝氨酸蛋白酶
紫外分光光度法
proteinaseK
activity detection
serine protease
ultraviolet spectrophotometry
