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嗜盐古菌地中海富盐菌中硝酸盐同化基因的鉴定与转录分析。

Identification and transcriptional analysis of nitrate assimilation genes in the halophilic archaeon Haloferax mediterranei.

作者信息

Lledó Belén, Marhuenda-Egea Frutos C, Martínez-Espinosa Rosa María, Bonete María José

机构信息

División de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de Alicante, Ap. 99, E- 03080, Alicante, Spain.

出版信息

Gene. 2005 Nov 21;361:80-8. doi: 10.1016/j.gene.2005.07.011. Epub 2005 Sep 22.

Abstract

Sequencing a 6,720-bp segment of the extreme halophilic archaeon Haloferax mediterranei genome has revealed the genomic organization of the putative structural genes for nitrate assimilation. We report a gene operon containing nasABC and nasD gene. nasA encodes an assimilatory nitrate reductase, nasB codes for a membrane protein with similarity to the NarK transporter, nasC encodes a protein with similarity to MobA; and nasD codes for an assimilatory ferredoxin-dependent nitrite reductase. Reverse transcription-PCR and primer extension experiments have demonstrated the existence of one polycistronic messenger nasABC and one monocistronic nasD initiated from a different promoter. The gene order and the grouping in two adjacent transcriptional units constitutes a novel organization of nas genes. The promoter regions harbor direct palindromes reminiscent of target sites for binding of a hypotetical regulatory protein(s). Transcription of the nasABC and nasD regions was found to be repressed by the presence of ammonium as nitrogen source.

摘要

对极端嗜盐古菌地中海嗜盐栖热菌(Haloferax mediterranei)基因组的一段6720 bp片段进行测序,揭示了假定的硝酸盐同化结构基因的基因组组织。我们报道了一个包含nasABC和nasD基因的基因操纵子。nasA编码同化型硝酸还原酶,nasB编码一种与NarK转运蛋白相似的膜蛋白,nasC编码一种与MobA相似的蛋白;nasD编码一种同化型铁氧化还原蛋白依赖性亚硝酸还原酶。逆转录PCR和引物延伸实验证明存在一个多顺反子信使nasABC和一个从不同启动子起始的单顺反子nasD。基因顺序以及在两个相邻转录单元中的分组构成了nas基因的一种新组织形式。启动子区域含有直接回文序列,让人联想到假定的调节蛋白结合的靶位点。发现以铵作为氮源时,nasABC和nasD区域的转录受到抑制。

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