Matsuda M, Kuribayashi T, Yamamoto S, Millar B C, Moore J E
Laboratory of Molecular Biology, Graduate School of Environmental Health Sciences, Azabu University, Sagamihara, 252-5201, Japan.
Department of Bacteriology, Northern Ireland Public Health Laboratory, Belfast City Hospital, Belfast, BT9 7AD, Northern Ireland, UK.
Folia Microbiol (Praha). 2016 Jan;61(1):57-62. doi: 10.1007/s12223-015-0405-z. Epub 2015 Jun 30.
An arsenate susceptibility test was performed with transformed and cultured Escherichia coli DH5α cells, which carried recombinant DNA of full-length arsenic (ars) operon, namely a putative membrane permease, ArsP; a transcriptional repressor, ArsR; an arsenate reductase, ArsC; and an arsenical-resistance membrane transporter, Acr3, from the Japanese urease-positive thermophilic Campylobacter lari (UPTC) CF89-12. The E. coli DH5α transformant showed reduced susceptibility to arsenate (1536 μg/mL), compared to the control. Thus, these ars four-genes from the UPTC CF89-12 strain cells could confer a reduced susceptibility to arsenate in the transformed and E. coli DH5α cells. E. coli transformants with truncated ars operons, acr3 (acr3) and arsC-acr3 (∆arsC-acr3), of the ars operon, showed an MIC value of 384 μg/mL (384 μg/mL), similar to the E. coli cells which carried the pGEM-T vector (control). Reverse transcription PCR confirmed in vivo transcription of recombinant full-length ars operon and deletion variants (∆acr3 and ∆arsC-acr3) in the transformed E. coli cells.
对携带来自日本脲酶阳性嗜热弯曲菌（UPTC）CF89 - 12的全长砷（ars）操纵子重组DNA的转化培养大肠杆菌DH5α细胞进行了砷酸盐敏感性测试。该重组DNA包含一个假定的膜通透酶ArsP、一个转录阻遏物ArsR、一个砷酸盐还原酶ArsC和一个抗砷膜转运蛋白Acr3。与对照相比，大肠杆菌DH5α转化体对砷酸盐的敏感性降低（1536μg/mL）。因此，来自UPTC CF89 - 12菌株细胞的这四个ars基因可使转化的大肠杆菌DH5α细胞对砷酸盐的敏感性降低。携带ars操纵子截短体acr3（acr3）和arsC - acr3（∆arsC - acr3）的大肠杆菌转化体的最低抑菌浓度（MIC）值为384μg/mL（384μg/mL），与携带pGEM - T载体的大肠杆菌细胞（对照）相似。逆转录PCR证实了重组全长ars操纵子及其缺失变体（∆acr3和∆arsC - acr3）在转化的大肠杆菌细胞中的体内转录。
Zotero 插件