Synthesis of membrane proteins in eukaryotic cell-free systems

被引:48
|
作者
Sachse, Rita
Wuestenhagen, Doreen
Samalikova, Maria
Gerrits, Michael [2 ]
Bier, Frank F. [3 ]
Kubick, Stefan [1 ]
机构
[1] Fraunhofer Inst Biomed Engn IBMT, Branch Potsdam Golm, D-14476 Potsdam, Germany
[2] RiNA GmbH, Berlin, Germany
[3] Univ Potsdam, Inst Biochem & Biol, Potsdam, Germany
来源
ENGINEERING IN LIFE SCIENCES | 2013年 / 13卷 / 01期
关键词
Cell-free protein expression; In vitro protein synthesis; Labeled membrane proteins; Synthetic glycoprotein; POST-TRANSLATIONAL MODIFICATION; IN-VITRO TRANSLATION; MESSENGER-RNA; N-GLYCOSYLATION; EXPRESSION; RECEPTOR; PHOSPHORYLATION; GAMMA; IDENTIFICATION; TRANSLOCATION;
D O I
10.1002/elsc.201100235
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Cell-free protein synthesis (CFPS) is a valuable method for the fast expression of difficult-to-express proteins as well as posttranslationally modified proteins. Since cell-free systems circumvent possible cytotoxic effects caused by protein overexpression in living cells, they significantly enlarge the scale and variety of proteins that can be characterized. We demonstrate the high potential of eukaryotic CFPS to express various types of membrane proteins covering a broad range of structurally and functionally diverse proteins. Our eukaryotic cell-free translation systems are capable to provide high molecular weight membrane proteins, fluorescent-labeled membrane proteins, as well as posttranslationally modified proteins for further downstream analysis.
引用
收藏
页码:39 / 48
页数:10
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