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首页> 外文期刊>Journal of bacteriology >Cloning, DNA sequence, and complementation analysis of the Salmonella typhimurium hemN gene encoding a putative oxygen-independent coproporphyrinogen III oxidase.
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Cloning, DNA sequence, and complementation analysis of the Salmonella typhimurium hemN gene encoding a putative oxygen-independent coproporphyrinogen III oxidase.

机译：鼠伤寒沙门氏菌hemN基因的克隆，DNA序列和互补分析，该基因编码一种与氧无关的拟卟啉原III氧化酶。

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摘要

Coproporphyrinogen oxidation is a last step in heme biosynthesis. The biochemically characterized eukaryotic coproporphyrinogen III oxidases have an obligate requirement for molecular oxygen, and a similar enzyme is encoded by the hemF gene in Salmonella typhimurium. Anaerobic heme synthesis requires an oxygen-independent coproporphyrinogen oxidase, which is probably encoded by the hemN gene in S. typhimurium. The hemN gene has been cloned from an insertion mutant. The nucleotide sequence was obtained and used for PCR amplification of the wild-type gene. A single open reading frame was identified as the hemN gene on the basis of its interruption by the insertion mutation and plasmid complementation studies of hemF hemN double mutants. The predicted HemN protein has 38% amino acid sequence identity to a putative anaerobic Rhodobacter sphaeroides coproporphyrinogen oxidase. The hemN RNA 5' end and the inferred transcription initiation site were mapped by primer extension. The 52.8-kDa HemN protein is expressed from the second ATG codon of the hemN open reading frame. An open reading frame with an unknown function directly upstream of hemN has a striking amino acid sequence, including 11 acidic residues in a row.
机译：卟啉原原的氧化是血红素生物合成的最后一步。具有生化特征的真核原卟啉原III氧化酶对分子氧有严格的要求，而鼠伤寒沙门氏菌中的hemF基因则编码类似的酶。厌氧血红素合成需要不依赖氧的共原卟啉原氧化酶，其可能由鼠伤寒沙门氏菌中的hemN基因编码。 hemN基因已从插入突变体克隆。获得了核苷酸序列，并将其用于野生型基因的PCR扩增。基于hemF hemN双突变体的插入突变和质粒互补研究，基于其中断，将单个开放阅读框鉴定为hemN基因。预测的HemN蛋白与推定的厌氧球形红细菌共卟啉原氧化酶具有38％的氨基酸序列同一性。 hemN RNA 5'末端和推断的转录起始位点通过引物延伸进行定位。从hemN开放阅读框的第二个ATG密码子表达52.8kDa的HemN蛋白。在hemN上游直接具有未知功能的开放阅读框具有引人注目的氨基酸序列，包括连续11个酸性残基。

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