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在弯曲杆菌中砷四基因操纵子的分子鉴定

Molecular identification of an arsenic four-gene operon in Campylobacter lari.

机构信息

Laboratory of Molecular Biology, Graduate School of Environmental Health Sciences, Azabu University, Sagamihara 252-5201, Japan.

出版信息

Folia Microbiol (Praha). 2013 May;58(3):253-60. doi: 10.1007/s12223-012-0207-5. Epub 2012 Nov 7.

Abstract

An arsenic (ars) four-gene operon, containing genes encoding a putative membrane permease (ArsP), a transcriptional repressor (ArsR), an arsenate reductase (ArsC) and an arsenical-resistance membrane transporter (Acr3) was first identified in urease-positive thermophilic Campylobacter (UPTC) isolate, CF89-12. UPTC CF89-12 and some other Campylobacter lari isolates contained their ars four-genes, similarly, differing from that in the reference C. lari RM2100 strain. Two putative promoters and a putative terminator were identified for the operon in UPTC CF89-12. In vivo transcription of the operon was confirmed in the UPTC cells. PCR experiments using two primer pairs designed in silico to amplify two arsR and arsC-acr3 segments, respectively, generated two amplicons, approximately 200 and 350 base pairs, with all 31 of 31 and 19 of 31 C. lari isolates (n = 17 for UPTC; n = 14 for UN C. lari), respectively. An inverted repeat forming a dyad structure, a potential binding site for a transcriptional repressor, was identified in the promoter region. Within the deduced 61 amino acids sequence of the putative arsR open reading frame from the UPTC CF89-12, a metal binding box and a DNA-binding helix-turn-helix motif were identified. The UPTC CF89-12 and some other UPTC isolates isolated from natural environment were resistant to arsenate.

摘要

砷(ars)四基因操纵子,包含编码假定膜透性酶(ArsP)、转录抑制剂(ArsR)、砷酸盐还原酶(ArsC)和砷抗性膜转运蛋白(Acr3)的基因,最初在脲酶阳性嗜热弯曲杆菌(UPTC)分离株 CF89-12 中被鉴定。UPTC CF89-12 和其他一些 Lari 弯曲杆菌分离株含有它们的 ars 四基因,与参考菌株 C. lari RM2100 不同。在 UPTC CF89-12 中,该操纵子鉴定出两个假定启动子和一个假定终止子。在 UPTC 细胞中证实了操纵子的体内转录。使用两个引物对设计的 PCR 实验,分别扩增两个 arsR 和 arsC-acr3 片段,产生了两个约 200 和 350 碱基对的扩增子,所有 31 个弯曲杆菌分离株(n = 17 UPTC;n = 14 UN C. lari)都得到了扩增。在启动子区域鉴定到一个形成二联体结构的反向重复,这是转录抑制剂的潜在结合位点。在 UPTC CF89-12 的假定 arsR 开放阅读框的推导 61 个氨基酸序列中,鉴定到一个金属结合盒和一个 DNA 结合螺旋-转角-螺旋基序。从自然环境中分离到的 UPTC CF89-12 和其他一些 UPTC 分离株对砷酸盐具有抗性。

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