Scott D L, Ramanathan S, Shi W, Rosen B P, Daunert S
Department of Chemistry, University of Kentucky, Lexington 40506-0055, USA.
Anal Chem. 1997 Jan 1;69(1):16-20. doi: 10.1021/ac960788x.
A bacterial sensing system that responds selectively to antimonite and arsenite has been investigated. The bacteria used in these studies have been genetically engineered to produce the enzyme beta-galactosidase in response to these ions. This is accomplished by using a plasmid that incorporates the gene for beta-galactosidase (reporter gene) under the control of the promoter of the ars operon. This plasmid also encodes for the ArsR protein, a regulatory protein of the ars operon, which, in the absence of antimonite or arsenite, restricts the expression of beta-galactosidase. In the presence of antimonite or arsenite the ArsR protein is released from the operator/ promoter region of the ars operon and beta-galactosidase is expressed. The activity of this enzyme was monitored electrochemically using p-aminophenyl beta-D-galactopyranoside as the substrate. The bacterial sensing system responds selectively to arsenite and antimonite (and to a lesser extent arsenate) and shows no significant response to phosphate, sulfate, nitrate, and carbonate.
一种对亚锑酸盐和亚砷酸盐具有选择性响应的细菌传感系统已被研究。这些研究中使用的细菌经过基因工程改造,可响应这些离子产生β-半乳糖苷酶。这是通过使用一种质粒来实现的,该质粒在ars操纵子启动子的控制下整合了β-半乳糖苷酶基因(报告基因)。该质粒还编码ArsR蛋白,它是ars操纵子的一种调节蛋白,在没有亚锑酸盐或亚砷酸盐的情况下,会限制β-半乳糖苷酶的表达。在有亚锑酸盐或亚砷酸盐存在时,ArsR蛋白从ars操纵子的操纵子/启动子区域释放,β-半乳糖苷酶得以表达。使用对氨基苯基β-D-吡喃半乳糖苷作为底物,通过电化学方法监测该酶的活性。该细菌传感系统对亚砷酸盐和亚锑酸盐(以及在较小程度上对砷酸盐)具有选择性响应,而对磷酸盐、硫酸盐、硝酸盐和碳酸盐无明显响应。