Purification of the recombinant green fluorescent protein from tobacco plants using alcohol/salt aqueous two-phase system and hydrophobic interaction chromatography
Purification of the recombinant green fluorescent protein from tobacco plants using alcohol/salt aqueous two-phase system and hydrophobic interaction chromatography
复制标题
使用醇/盐水两相系统和疏水相互作用层析纯化烟草植物中的重组绿色荧光蛋白
DOI：
10.1186/s12896-019-0590-y
复制
发表时间：
2019-12-09
影响因子：
3.5
通讯作者：
Wang, Sheng
中科院分区：
文献类型：
--
作者：
Dong, Jie;Ding, Xiangzhen;Wang, Sheng
BackgroundThe green fluorescent protein (GFP) has been regarded as a valuable tool and widely applied as a biomarker in medical applications and diagnostics. A cost-efficient upstream expression system and an inexpensive downstream purification process will meet the demands of the GFP protein with high-purity.ResultsThe recombinant GFP was transiently expressed in an active form in agoinoculatedNicotiana benthamianaleaves by usingTobacco mosaic virus(TMV) RNA-based overexpression vector (TRBO). The yield of recombinant GFP was up to ~ 60% of total soluble proteins (TSP). Purification of recombinant GFP from the clarified lysate ofN. benthanianaleaves was achieved by using an alcohol/salt aqueous two-phase system (ATPS) and following with a further hydrophobic interaction chromatography (HIC). The purification process takes only ~ 4 h and can recover 34.1% of the protein. The purity of purified GFP was more than 95% and there were no changes in its spectroscopic characteristics.ConclusionsThe strategy described here combines the advantages of both the economy and efficiency of plant virus-based expression platform and the simplicity and rapidity of environmentally friendly alcohol/salt ATPS. It has a considerable potential for the development of a cost-efficient alternative for production of recombinant GFP.
