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。
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.
出处
《河北农业大学学报》
CAS
CSCD
北大核心
2016年第2期15-20,共6页
Journal of Hebei Agricultural University
基金
河北省普通高等学校高层次人才科学研究项目(GCC2014029)
青年拔尖人才项目(BJ2014040)部分资助
关键词
拟南芥
NF-YC9
重组蛋白
工程菌
MBP
亲和纯化
Arabidopsis thaliana
NF-YC9
recombinant protein
engineering bacteria
MBP
affinity purification
