Purification of green fluorescent protein using a two-intein system

doi:10.1007/s00253-007-1233-0

	Purification of green fluorescent protein using a two-intein system
	Zhao, Zhonglin 2; Lu, Wei 1; Dun, Baoqing 3; Jin, Dan 4; Ping, Shuzhen 1; Zhang, Wei 1; Chen, Ming 1; Xu, Ming-Qun 5; Lin, Min 1
刊名	APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
	2008
卷号	77 期号:5 页码:1175-1180
关键词	intein protein splicing green fluorescent protein protein purification
ISSN号	0175-7598
DOI	10.1007/s00253-007-1233-0
通讯作者	Zhao, Zhonglin
英文摘要	A two-intein purification system was developed for the affinity purification of GFPmut3, a mutant of green fluorescent protein. The GFPmut3 was sandwiched between two self-cleaving inteins. This approach avoided the loss of the target protein which may result from in vivo cleavage of a single intein tag. The presence of N- and C-terminal chitin-binding domains allowed the affinity purification by a single-affinity chitin column. After the fusion protein was expressed and immobilized on the affinity column, self-cleavage of the inteins was sequentially induced to release the GFPmut3. The yield was 2.41 mg from 1 l of bacterial culture. Assays revealed that the purity was up to 98% of the total protein. The fluorescence and circular dichroism spectrum of GFPmut3 demonstrated that the purified protein retains the correctly folded structure and function.
学科主题	Biotechnology & Applied Microbiology ; BIOTECHNOLOGY & APPLIED MICROBIOLOGY
语种	英语
出版者	SPRINGER
WOS记录号	WOS:000251606900022
内容类型	期刊论文
源URL	[http://111.203.20.206/handle/2HMLN22E/5636]
专题	作物科学研究所_遗传育种系
作者单位	1.Chinese Acad Agr Sci, Biotechnol Res Inst, Beijing 100081, Peoples R China 2.China Agr Univ, Coll Biol Sci, Beijing 100094, Peoples R China 3.Chinese Acad Agr Sci, Inst Crop Sci, Beijing 100081, Peoples R China 4.SW Univ, Biotechnol Res Ctr, Chongqing 400716, Peoples R China 5.New England Biolabs Inc, Beverly, MA 01915 USA
推荐引用方式 GB/T 7714	Zhao, Zhonglin,Lu, Wei,Dun, Baoqing,et al. Purification of green fluorescent protein using a two-intein system[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2008,77(5):1175-1180.
APA	Zhao, Zhonglin.,Lu, Wei.,Dun, Baoqing.,Jin, Dan.,Ping, Shuzhen.,...&Lin, Min.(2008).Purification of green fluorescent protein using a two-intein system.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,77(5),1175-1180.
MLA	Zhao, Zhonglin,et al."Purification of green fluorescent protein using a two-intein system".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 77.5(2008):1175-1180.