摘要
FT基因是植物成花素,在植物的开花调控中起着重要作用。构建了用于原核表达的FT-eGFP表达载体,并在大肠杆菌BL21中进行重组蛋白的诱导表达。从IPTG诱导浓度、诱导时间和诱导温度等方面进行了细致的分析,最终建立了FT-eGFP融合蛋白诱导表达的优化体系:当菌液OD600=0.6-1.0时,采用IPTG1.0mol/L,在28℃诱导表达6h。摸索和建立了利用HisTrapKit标签,经过镍柱纯化,纯化目的蛋白的技术体系,为进一步研究FT基因在植物开花调控中的应用奠定了基础。
FT is a key gene in regulating the flowering time of Arabidopsis. In this research,expression vector of FT-eGFP in prokaryotic cells was constructed and the fused protein expression was induced in E. coli BL21. IPTG concentration,induction time and induction temperature were determined to set up optimal condition for fused protein expression purification. When OD600 of E. coli liqulid culture was 0. 6 - 1.0,final concentration of 1.0 mol/L IPTG was applied into the culture to induce fused protein followed by incubation at 28℃ for 6 hours. FT-eGFP fuse protein was then purified with His Trap Kit. This research makes it possible to investigate the function of FT protein in promoting plant flowering.
出处
《生物技术通报》
CAS
CSCD
北大核心
2010年第3期99-103,共5页
Biotechnology Bulletin
基金
国家"863"项目(2008AA10Z1562)
北京市园林绿化局菊花育种创新平台(YlHH2008002)
