| 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。 |
| 关键词: 磁小体 固相载体 碱性磷酸酶 磁性免疫ELISA 趋磁细菌 |
| DOI：10.11841/j.issn.1007-4333.2018.04.02 |
| 投稿时间：2017-05-09 |
| 基金项目: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 |
|
| Analysis of alkaline phosphatase activity of magnetosome |
|
XU Junjie1,2, WANG Manman1, LIU Lingzi2, LI Feng1, TIAN Jiesheng2
|
| (1.College of Life Science, Huaibei normal university, Huaibei 235000, China;2.College of Biological Sciences/State Key Laboratory of Agricultural Biotechnology, China Agricultural University, Beijing 100193, China) |
| Abstract: |
| 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 location 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. |
| Key words: magnetosome solid phase carrier alkaline phosphatase magnetic immune ELISA magnetotactic bacteria |
