含有培养的人类细胞提取物的无细胞蛋白质合成系统。

Cell-free protein synthesis systems with extracts from cultured human cells.

作者信息

Mikami Satoshi, Kobayashi Tominari, Imataka Hiroaki

机构信息

RIKEN Systems and Structural Biology Center, Yokohama, Japan.

出版信息

Methods Mol Biol. 2010;607:43-52. doi: 10.1007/978-1-60327-331-2_5.

DOI:10.1007/978-1-60327-331-2_5

PMID:20204847

Abstract

Cell-free protein synthesis systems have been established with extracts from cultured human cells, HeLa, and hybridoma cells. The former cell line is used to prepare extracts for robust translation, whereas the extract from the latter is primarily employed for expression of glycoproteins. Productivity of both systems can be enhanced by addition of K3L and GADD34, factors that diminish phosphorylation of eIF2alpha. The coupled transcription/translation system is also available as a convenient tool, particularly for the production of large recombinant proteins.

摘要

已利用培养的人类细胞、HeLa细胞和杂交瘤细胞的提取物建立了无细胞蛋白质合成系统。前一种细胞系用于制备用于高效翻译的提取物，而后一种细胞的提取物主要用于糖蛋白的表达。通过添加K3L和GADD34（可减少eIF2α磷酸化的因子），这两种系统的生产力均可得到提高。耦合转录/翻译系统也是一种方便的工具，尤其适用于大型重组蛋白的生产。