Cai Lin, Rensing Christopher, Li Xiangyang, Wang Gejiao
Huazhong Agricultural University, Wuhan, China.
Appl Microbiol Biotechnol. 2009 Jun;83(4):715-25. doi: 10.1007/s00253-009-1929-4. Epub 2009 Mar 13.
This study describes three gene clusters involved in arsenic redox transformation of two arsenite oxidizers: Achromobacter sp. SY8 and Pseudomonas sp. TS44. A 17.5-kb sequence containing the arsenite oxidase (aox) gene cluster (aoxX-aoxS-aoxR and aoxA-aoxB-aoxC-aoxD) was isolated from SY8 using a fosmid library approach. Similarly, a 14.6-kb sequence including the aox cluster (arsD-arsA-aoxA-aoxB) and the arsenic resistance (ars) gene cluster (arsC1-arsR-arsC2-ACR3-arsH-dual specificity phosphatase (DSP)-glyceraldehyde-3-phosphate dehydrogenase (GAPDH)-major facilitator superfamily (MFS)) was obtained from TS44 by inverse polymerase chain reaction (PCR). According to reverse transcription (RT) PCR experiments, SY8 aoxXSR and aoxABCD transcribed as two different transcripts in opposite directions, and TS44 aox and ars clusters transcribed as a single transcript in their respective cluster. All of these genes were found to be upregulated by the addition of arsenite [As(III)], arsenate [As(V)], and antimonite [Sb(III)], except that TS44 arsC1-arsR appeared to be expressed constitutively. The SY8 aox cluster was predicted to be regulated by a two-component signal transduction system and a potential regulatory model was proposed. The TS44 aox cluster is unusual since it contains structural genes only and arsDA in its upstream. The TS44 ars cluster includes several genes previously identified not associated with arsenic resistance or transformation. This study showed novel structures and arrangements of arsenic gene clusters associated with bacterial As(III) oxidation and As(V) reduction.
本研究描述了参与两种亚砷酸盐氧化菌(无色杆菌属SY8菌株和假单胞菌属TS44菌株)砷氧化还原转化的三个基因簇。采用fosmid文库法从SY8菌株中分离出一个包含亚砷酸盐氧化酶(aox)基因簇(aoxX-aoxS-aoxR和aoxA-aoxB-aoxC-aoxD)的17.5 kb序列。同样,通过反向聚合酶链反应(PCR)从TS44菌株中获得了一个14.6 kb的序列,该序列包括aox基因簇(arsD-arsA-aoxA-aoxB)和抗砷(ars)基因簇(arsC1-arsR-arsC2-ACR3-arsH-双特异性磷酸酶(DSP)-甘油醛-3-磷酸脱氢酶(GAPDH)-主要促进剂超家族(MFS))。根据逆转录(RT)PCR实验,SY8菌株的aoxXSR和aoxABCD以相反方向转录为两种不同的转录本,TS44菌株的aox和ars基因簇在各自的基因簇中作为单一转录本转录。除TS44菌株的arsC1-arsR似乎组成性表达外,所有这些基因在添加亚砷酸盐[As(III)]、砷酸盐[As(V)]和亚锑酸盐[Sb(III)]后均被上调。预测SY8菌株的aox基因簇受双组分信号转导系统调控,并提出了一个潜在的调控模型。TS44菌株的aox基因簇不同寻常,因为它仅包含结构基因且在其上游有arsDA。TS44菌株的ars基因簇包括几个先前鉴定出的与抗砷或砷转化无关的基因。本研究展示了与细菌As(III)氧化和As(V)还原相关的砷基因簇的新结构和排列。