Heterologous expression of clostridial hydrogenase in the cyanobacterium Synechococcus PCC7942

被引：58

作者：

Asada, Y

Koike, Y

Schnackenberg, J

Miyake, M

Uemura, I

Miyake, J

机构：

[1] Res Inst Innovat Technol Earth, Tsukuba, Ibaraki 3058566, Japan

[2] Ind Technol Ctr, Okayama 7011296, Japan

[3] Tokyo Gas Co Ltd, Frontier Technol Res Inst, Yokohama, Kanagawa 2300045, Japan

[4] Natl Inst Adv Interdisciplinary Res, Tsukuba, Ibaraki 3058562, Japan

[5] Agcy Ind Sci & Technol, Natl Inst Biosci & Human Technol, Tsukuba, Ibaraki 3058566, Japan

来源：

BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION | 2000年 / 1490卷 / 03期

关键词：

Synechococcus PCC 7942; heterologous gene expression; hydrogen production;

D O I：

10.1016/S0167-4781(00)00010-5

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The Clostridium pasteurianum hydrogenase I has been expressed in the cyanobacterium Synechococcus PCC7942. The Shine-Dalgarno sequence of the structural gene encoding hydrogenase I from C. pasteurianum was changed to that of the cat (chloramphenicol acetyltransferase) gene. The hydrogenase gene was cloned downstream of a strong promoter, isolated from Synechococcus PCC7942, with the cat gene as a reporter gene. Expression of clostridial hydrogenase was confirmed by Western and Northern blot analyses in Synechococcus and Escherichia coli, whereas in vivo/in vitro measurements and activity staining of soluble proteins separated on non-denaturing polyacrylamide gels revealed functional expression of hydrogenase only in cyanobacterial cells. The changed Shine-Dalgarno sequence appeared to be essential for the functional expression of clostridial hydrogenase in Synechococcus, but had no influence on the expression and activity of clostridial hydrogenase expressed in E. coli. (C) 2000 Elsevier Science B.V. All rights reserved.

引用

页码：269 / 278

页数：10

共 49 条

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