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对编码荚膜红细菌7铁铁氧还蛋白(FdII)的fdxA基因进行转录分析和启动子定位。

Transcriptional analysis and promoter mapping of the fdxA gene which encodes the 7Fe ferredoxin (FdII) of Rhodobacter capsulatus.

作者信息

Duport C, Jouanneau Y, Vignais P M

机构信息

Biochimie Microbienne (UA 1130 CNRS alliée à l'INSERM), Département de Biologie Moléculaire et Structurale, Grenoble, France.

出版信息

Mol Gen Genet. 1992 Jan;231(2):323-8. doi: 10.1007/BF00279806.

Abstract

The structural gene (fdxA) coding for ferredoxin II (FdII) of the photosynthetic bacterium Rhodobacter capsulatus has been previously cloned and sequenced. Transcription of the fdxA gene was studied by mRNA analyses and by use of plasmid-borne fdxA::lacZ translational fusions. The transcription start site was mapped 23 bp upstream of the initiation codon, as deduced from analysis of mRNA by mung bean nuclease protection and primer extension experiments. A motif resembling the canonical sequence observed in sigma 70-dependent Escherichia coli promoters is present at the expected distance (-10/-35) from the proposed transcription start site. mRNA analysis by Northern hybridization revealed a fdxA-specific transcript of approximately 0.4 kb, indicating that fdxA is transcribed as a single gene.fdxA expression, as measured by the activity of the fdxA::lacZ fusion, was found to be constant during growth and reached a similar level under all growth conditions tested. These results suggest that FdII is constitutively synthesized in R. capsulatus.

摘要

光合细菌荚膜红细菌(Rhodobacter capsulatus)中编码铁氧化还原蛋白II(FdII)的结构基因(fdxA)此前已被克隆和测序。通过mRNA分析以及使用质粒携带的fdxA::lacZ翻译融合体对fdxA基因的转录进行了研究。通过绿豆核酸酶保护和引物延伸实验对mRNA进行分析,推断转录起始位点位于起始密码子上游23 bp处。在距推测的转录起始位点预期距离(-10 / -35)处存在一个类似于在依赖σ70的大肠杆菌启动子中观察到的典型序列的基序。通过Northern杂交进行的mRNA分析揭示了一个约0.4 kb的fdxA特异性转录本,表明fdxA作为单个基因进行转录。通过fdxA::lacZ融合体的活性测定发现,fdxA表达在生长过程中保持恒定,并且在所有测试的生长条件下达到相似水平。这些结果表明,FdII在荚膜红细菌中是组成型合成的。

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