Isolation of several anti-stress genes from the halotolerant green alga Chlamydomonas by simple functional expression screening with Escherichia coli

被引:29
作者
Miyasaka, H
Kanaboshi, H
Ikeda, K
机构
[1] Kansai Elect Power Co, Tech Res Ctr, Amagasaki, Hyogo 6610974, Japan
[2] Kansai Environm Engn Ctr Co Ltd, Chuo Ku, Osaka 5410052, Japan
关键词
anti-stress gene; ascorbate peroxidase; bbc1; Chlamydomonas; glutathione peroxidase;
D O I
10.1023/A:1008982332139
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
To isolate anti-stress genes from a halotolerant marine green alga, a simple screening method based on the bacterial expression system was examined. The method consisted of three steps: (i) directional cDNA library construction in a lambda ZAPII vector, (ii) in vivo mass excision of a lambda ZAPII library into phagemid DNA, and (iii) screening for anti-salt-stress and anti-oxidative-stress genes by culturing bacterial cells carrying the in vivo excised phagemid on selection agar plates with a high concentration of NaCl and/or methyl viologen (MV), and by isolating stress-tolerant bacterial colonies. By this method, we screened the cDNA library of halotolerant Chlamydomonas sp. strain W-80, and isolated several stress-related gene homologs, such as glutathione peroxidase, ascorbate peroxidase, bbc1 (breast basic conserved; a low temperature induced gene in Brassica napus), and cyanide-resistant alternative oxidase.
引用
收藏
页码:23 / 29
页数:7
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