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Transcriptional Organization and Regulation of Magnetosome Operons in Magnetospirillum gryphiswaldense

机译:灰螺螺螺旋藻中核小体操纵子的转录组织和调控

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摘要

Genes involved in magnetite biomineralization are clustered within the genomic magnetosome island of Magnetospirillum gryphiswaldense. Their transcriptional organization and regulation were studied by several approaches. Cotranscription of genes within the mamAB, mamDC, and mms clusters was demonstrated by reverse transcription-PCR (RT-PCR) of intergenic regions, indicating the presence of long polycistronic transcripts extending over more than 16 kb. The transcription start points of the mamAB, mamDC, and mms operons were mapped at 22 bp, 52 bp, and 58 bp upstream of the first genes of the operons, respectively. Identified −10 and −35 boxes of the PmamAB, PmamDC, and Pmms promoters showed high similarity to the canonical σ70 recognition sequence. The transcription of magnetosome genes was further studied in response to iron and oxygen. Transcripts of magnetosome genes were detected by RT-PCR both in magnetic cells grown microaerobically under iron-sufficient conditions and in nonmagnetic cells grown either aerobically or with iron limitation. The presence of transcripts was found to be independent of the growth phase. Further results from partial RNA microarrays targeting the putative magnetosome transcriptome of M. gryphiswaldense and real-time RT-PCR experiments indicated differences in expression levels depending on growth conditions. The expression of the mam and mms genes was down-regulated in nonmagnetic cells under iron limitation and, to a lesser extent, during aerobic growth compared to that in magnetite-forming cells grown microaerobically under iron-sufficient conditions.
机译:磁铁矿生物矿化中涉及的基因聚集在Magnetospirillum gryphiswaldense的基因组磁小体岛内。通过几种方法研究了它们的转录组织和调控。 mamAB,mamDC和mms簇内基因的共转录通过基因间区域的逆转录PCR(RT-PCR)证明,表明存在超过16 kb的长多顺反子转录本。 mamAB,mamDC和mms操纵子的转录起始点分别位于操纵子第一个基因上游的22 bp,52 bp和58 bp。鉴定出的PmamAB,PmamDC和Pmms启动子的-10和-35盒与标准σ 70 识别序列具有高度相似性。磁小体基因的转录进一步研究了对铁和氧的响应。通过RT-PCR,在铁充足的条件下以微需氧方式生长的磁性细胞和需氧或铁受限的非磁性细胞中,通过RT-PCR检测了磁小体基因的转录本。发现转录本的存在与生长期无关。来自针对假定的M. gryphiswaldense磁小体转录组的部分RNA微阵列的进一步结果和实时RT-PCR实验表明,表达水平的差异取决于生长条件。 mam和mms基因的表达在铁受限的非磁性细胞中下调,并且与在铁充足的条件下微厌氧生长的磁铁矿形成细胞相比,有氧生长过程中的表达程度较低。

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