英特因
蛋白质剪接
蛋白质标签
化学
RNA剪接
残留物(化学)
反式剪接
肽
天然化学连接
麦芽糖结合蛋白
生物化学
融合蛋白
立体化学
化学合成
重组DNA
基因
体外
核糖核酸
作者
Belinda M. Lew,Kenneth V. Mills,Henry Paulus
出处
期刊:Biopolymers
[Wiley]
日期:1999-01-01
卷期号:51 (5): 355-362
被引量:27
标识
DOI:10.1002/(sici)1097-0282(1999)51:5<355::aid-bip5>3.0.co;2-m
摘要
Protein splicing in trans results in the ligation of two protein or peptide segments linked to appropriate intein fragments. We have characterized the trans-splicing reaction mediated by a naturally expressed, approximately 100-residue N-terminal fragment of the Mycobacterium tuberculosis intein and a synthetic peptide containing the 38 C-terminal intein residues, and found that the splicing reaction was very versatile and robust. The efficiency of splicing was nearly independent of temperature between 4 and 37°C and pH between 6.0 and 7.5, with only a slight decline at pH values as high as 8.5. In addition, there was considerable flexibility in the choice of the C-terminal intein fragment, no significant difference in protein ligation efficiency being observed between reactions utilizing the N-terminal fragment and either the naturally expressed 107-residue C-terminal portion of the intein, much smaller synthetic peptides, or the 107-residue C-terminal intein fragment modified by fusion of a maltose binding protein domain to its N-terminus. The ability to use different types of the C-terminal intein fragments and a broad range of reaction conditions make protein splicing in trans a versatile tool for protein ligation. © 2000 John Wiley & Sons, Inc. Biopoly 51: 355–362, 1999
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