关键词: 免疫沉淀, 磁珠, 蛋白标签, 微量蛋白

Abstract: In order to establish specific immunoprecipitation of few proteins in plant, GUS was used as a reporter gene which was marked with Flag and Myc tag on its N-terminal, and was transiently expressed in tobacco leaves via Agrobacterium-mediated method. Western blot analysis suggested the tagged GUS protein accumulated on a large scale in infiltrated tobacco leaves, which was helpful for specific purification of GUS protein. With anti- Flag M2 magnetic beads and anti-c-Myc agarose affinity gel as solid phase carrier, GUS protein was significantly enriched and purified after two-step immunoprecipitation. Western blot analysis indicated that enrichment efficiency of each step of our immunoprecipition was high. The results suggested the specific immunoprecipitation system could largely enrich the amount of tagged GUS protein. Moreover, GUS proteins were pure in final elution. This specific immunoprecipitation system with magnetic beads and protein tags would shed new lights on enrichment and purification of protein complexes of interest.

Key words: immunoprecipitation, magnetic beads, protein tags, few proteins