A LONG-LIVED BATCH REACTION SYSTEM OF CELL-FREE PROTEIN-SYNTHESIS

被引：50

作者：

KAWARASAKI, Y ^{[1
]}

KAWAI, T ^{[1
]}

NAKANO, H ^{[1
]}

YAMANE, T ^{[1
]}

机构：

[1] NAGOYA UNIV,SCH AGR,DEPT APPL BIOL SCI,CTR MOLEC BIOTECHNOL,CHIKUSA KU,NAGOYA,AICHI 464,JAPAN

来源：

ANALYTICAL BIOCHEMISTRY | 1995年 / 226卷 / 02期

关键词：

D O I：

10.1006/abio.1995.1231

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Several reaction conditions of cell-free protein synthesis such as temperatures, buffers, tRNAs, and creatine phosphate were intensively investigated and optimized to prolong protein synthesis and make it more efficiently in a batch system. As a result of these modifications, the protein synthesis reaction continued for 10 h so that about 30 mu g of dihydrofolate reductase (DHFR) protein derived from Escherichia coli was synthesized in 1 ml of reaction mixture. In this improved system, translational reactions of other mRNAs such as rabbit beta-globin, Xenopus beta-globin, and tobacco mosaic virus RNA also continued for about 10 h. In addition, protein synthesis directed by uncapped dhfr mRNA containing a viral cap-independent translation initiation-mediating sequence continued for 10 h, resulting in the synthesis of 18 mu g of DHFR protein per milliliter of reaction mixture. (C) 1995 Academic Press, Inc.

引用

页码：320 / 324

页数：5

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