Cai J, DuBow M S
Department of Microbiology and Immunology, McGill University, Montreal, Quebec, Canada.
Biodegradation. 1997;8(2):105-11. doi: 10.1023/a:1008281028594.
An arsenic oxyanion-inducible Escherichia coli chromosomal operon (arsRBC) has been previously identified. Construction of a luciferase transcriptional gene fusion (arsB::luxAB) showed that ars operon expression, plus concomitant cell luminescence, was inducible in a concentration-dependent manner by arsenic salts. The present study was conducted to evaluate the potential of the arsB::luxAB transcriptional gene fusion for use as a biosensor in monitoring the toxicity of arsenic compounds. Cultures from this gene fusion strain were exposed to increasing concentrations of the wood preservative chromated copper arsenate (CCA), as well as its constituents, sodium arsenate and chromated copper solution (CC). Analysis of luciferase activity revealed that the arsB::luxAB gene fusion was expressed in response to CCA and sodium arsenate, but not to the CC solution. The detection limit of arsenic was found to be 0.01 microgram As/ml (10 parts per billion, 10 ppb) and therefore well within the range of environmental concerns. A greater induction of luminescence by arsenate was observed when cells were limited for phosphate, as phosphate can act as a competitive inhibitor of arsenate ions. Our results suggest that the E. coli arsB::luxAB fusion strain has a promising future as a specific and sensitive biosensor for monitoring bioavailable levels and toxicity of arsenic near sites where CCA-treated wood has been used.
先前已鉴定出一种砷含氧阴离子诱导型大肠杆菌染色体操纵子(arsRBC)。构建荧光素酶转录基因融合体(arsB::luxAB)表明,ars操纵子的表达以及随之而来的细胞发光可被砷盐以浓度依赖的方式诱导。本研究旨在评估arsB::luxAB转录基因融合体作为生物传感器监测砷化合物毒性的潜力。将该基因融合菌株的培养物暴露于浓度不断增加的木材防腐剂铬酸铜砷(CCA)及其成分砷酸钠和铬酸铜溶液(CC)中。对荧光素酶活性的分析表明,arsB::luxAB基因融合体对CCA和砷酸钠有反应,但对CC溶液无反应。发现砷的检测限为0.01微克As/毫升(十亿分之十,10 ppb),因此完全在环境关注范围内。当细胞缺乏磷酸盐时,观察到砷酸盐对发光的诱导作用更大,因为磷酸盐可作为砷酸根离子的竞争性抑制剂。我们的结果表明,大肠杆菌arsB::luxAB融合菌株作为一种特异性和灵敏的生物传感器,在监测使用过CCA处理木材的地点附近砷的生物可利用水平和毒性方面具有广阔的前景。