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拟南芥NF-YC9重组蛋白的制备
Preparation of Arabidopsis NF-YC9 recombinant protein from the engineering bacteria
【摘要】 NF-YC9是模式植物拟南芥核因子Y家族成员之一,在开花调控等过程中发挥重要作用,但其蛋白的特性及其作用的分子机制尚待探究。本文首先扩增获得了NF-YC9基因的序列,采用重组技术构建了与麦芽糖结合蛋白MBP编码基因融合的重组表达载体,将其转化到大肠杆菌BL21中,进行了MBP-NF-YC9融合蛋白的诱导表达,并借助直链淀粉亲和层析介质纯化技术获得了电泳纯的MBP-NF-YC9重组蛋白,以备进一步对NF-YC9蛋白的生化特性进行全面检测。
【Abstract】 NF-YC9,as one member of NF-Y family,playes an important role in regulating flowering time of Arabidopsis.In this experiment,the gene sequence of NF-YC9 was amplified and cloned by RT-PCR,and then fused with the gene encoding maltose-binding protein(MBP)by using Gateway system.Then the plasmid p MBP-NF-YC9 was transformed into the engineering E.coli strain BL21.The optimum condition for the soluble MBP-NF-YC9 was established in the study.With the use of amylase-resin,the recombinant MBP-NF-YC9 proteins were purified and detected by SDS-PAGE.The purified recombinant protein will facilitate our further understanding about the biochemical characteristics of Arabidopsis NF-YC9.
【Key words】 Arabidopsis thaliana; NF-YC9; recombinant protein; engineering bacteria; MBP; affinity purification;
- 【文献出处】 河北农业大学学报 ,Journal of Agricultural University of Hebei , 编辑部邮箱 ,2016年02期
- 【分类号】Q946
- 【下载频次】127