磁小体的碱性磷酸酶活性分析  

Analysis of alkaline phosphatase activity of magnetosome

作　　者：徐俊杰[1,2] 王曼曼 刘凌子 李峰 田杰生[2] 

机构地区：[1]淮北师范大学生命科学学院,淮北235000 [2]中国农业大学生物学院/农业生物技术国家重点实验室,北京100193

出　　处：《中国农业大学学报》2018年第4期8-13,共6页Journal of China Agricultural University

基　　金：中国国家自然科学基金Grant No.31570037;农业生物技术国家重点实验室开放研究重点课题Grant No.2015SKLAB6-2

摘　　要：为测定磁小体的碱性磷酸酶活性及作为磁性免疫ELISA固相载体的可行性,设置不同显色时间、温度及pH条件,分析比较磁小体与商业化碱性磷酸酶的酶活。结果表明:商业化碱性磷酸酶显色时间较短,非常灵敏,但对温度和pH要求严格;微量的磁小体,需经过较长的显色时间,才能使显色剂显色,但温度和pH对磁小体的显色反应影响较小。通过不同的蛋白变性剂进行处理,表明磁小体的类碱性磷酸酶活性是其表面蛋白作用的结果。但磁小体微弱的显色能力,在磁性免疫ELISA检测时,没有显著影响。因此,磁小体可作为磁性免疫ELISA的固相载体,用于免疫检测。An experiment was carried out to determine alkaline phosphatase-like activity in magnetosomes and whether the magnetosomes could act as the solid carrier of magnetic immune ELISA. The activity of the magnetosome and the commercial alkaline phosphatase was compared under different coloring time, temperature and pH, The results showed that the time used for the color development of commercial alkaline phosphatase was very short and sensitive. However,the temperature and pH had certain limitations. The trace of the magnetosome required long time in color development, temperature and pH had little effect on the color reaction of magnetosome. Moreover, the Iooation of the magnetophilic alkaline phosphatase on magnetosome was preliminarily determined by the treatment of different protein denaturants. Based on these findings,it was concluded that the weak chromogenic ability of the magnetosome was not significantly affected when it was used for magnetic immunoassay ELISA. Therefore, the magnetosome could be used as a solid carrier for magnetic immune ELISA.

关 键 词：磁小体 固相载体 碱性磷酸酶 磁性免疫ELISA 趋磁细菌 

分 类 号：Q939.91[生物学—微生物学]