生物传感器在渔业监测中的研究进展

论述了生物传感器在监测渔业环境中的BOD、N、P、病原体、农药和重金属等方面的国内外最新进展 ,分析了生物传感器在海洋渔业应用中存在的主要问题及原因 ,同时阐述了利用基因工程微生物技术、分子烙印技术、多成分实时分析技术等先进技术提高生物传感器的性能并扩大其在海洋渔业领域中的应用范围 ,在海洋渔业环境中为实现生物传感器与计算机在线自动监测与控制 ,开拓新的广阔前景。     

微生物全细胞传感器在重金属生物可利用度监测中的研究进展

传统的物理化学方法主要用于测定环境重金属的总量,微生物全细胞传感器可以对土壤及水体环境的重金属生物可利用度进行监测.此外,微生物全细胞传感器还具有操作简单、快速、经济的特点,适用于污染事件的应急监测.微生物全细胞传感器的生物学元件主要由MerR、ArsR、RS等家族的金属调控蛋白和gfp、lux、luc等报告基因组成.调控蛋白、报告基因与微生物全细胞传感器的灵敏度、特异性和监测特点有关.受pH、金属螯合物及检测条件等因素的影响,不同的环境条件下的重金属生物可利用度是不同的.增加重金属在微生物细胞内的累积,进行调控蛋白的分子生物学改造,优化检测条件是提高传感器灵敏度、特异性和准确性的可行方案.实现污染物的原位和在线监测是微生物全细胞传感器的主要发展方向.     

Overexpression of NADH oxidase gene from Deinococcus geothermalis in Escherichia coli

When using stable enzyme genes from a thermophile to create a biosensor in Escherichia coli, it is vital that these genes be overexpressed in order to provide a sufficient supply of enzymes. In this study, overexpression of the NADH oxidase (Nox) gene from the thermophile Deinococcus geothermalis was successfully achieved with the aim of creating a stable biosensor active at room temperatures. To do so, modification of 10 nucleotides, GAAATTAACT, upstream of the start codon of the Nox gene was necessary.     

Overexpression of NADH oxidase gene from Deinococcus geothermalis in Escherichia coli

When using stable enzyme genes from a thermophile to create a biosensor in Escherichia coli, it is vital that these genes be overexpressed in order to provide a sufficient supply of enzymes. In this study, overexpression of the NADH oxidase (Nox) gene from the thermophile Deinococcus geothermalis was successfully achieved with the aim of creating a stable biosensor active at room temperatures. To do so, modification of 10 nucleotides, GAAATTAACT, upstream of the start codon of the Nox gene was necessary.     

Resistance and biosorption mechanism of silver ions by Bacillus cereus biomass

Biosorption of silver ions onto Bacillus cereus biomass was investigated. Overall kinetic experiments were performed for the determination of the necessary contact time for the attainment of equilibrium. It was found that the overall biosorption process was best described by pseudo second-order kinetic model. The crystals detected by scanning electron microscope and X-ray photoelectron spectroscopy suggested the precipitation was a possible mechanism of biosorption. The molecular genetics of silver resistance of B. cereus biomass was also detected and illustrated by a whole cell sensor tool.     

BOD生物传感器快速测定方法应用与探讨

介绍了微生物传感器快速测定生化需氧量(BOD)的方法,通过对标准溶液、工业废水、地表水、污水处理厂废水的检测,分析其精密度和准确度,通过与BOD_5的比对,对方法的存在问题及适用性进行了探讨和研究,为该方法的发展及其在环境监测中的实际应用提供了技术依据。     

Biological and chemical assessments of zinc ageing in field soils

As zinc (Zn) is both an essential trace element and potential toxicant, the effects of Zn fixation in soil are of practical significance. Soil samples from four field sites amended with ZnSO₄ were used to investigate ageing of soluble Zn under field conditions over a 2-year period. Lability of Zn measured using ⁶⁵Zn radioisotope dilution showed a significant decrease over time and hence evidence of Zn fixation in three of the four soils. However, 0.01 M CaCl₂ extractions and toxicity measurements using a genetically modified lux-marked bacterial biosensor did not indicate a decrease in soluble/bioavailable Zn over time. This was attributed to the strong regulatory effect of abiotic properties such as pH on these latter measurements. These results also showed that Zn ageing occurred immediately after Zn spiking, emphasising the need to incubate freshly spiked soils before ecotoxicity assessments.     

A GFP-based bacterial biosensor with chromosomally integrated sensing cassette for quantitative detection of Hg(II) in environment

A mercury biosensor was constructed by integrating biosensor genetic elements into E. coli JM109 chromosome in a single copy number, using the attP/attB recombination mechanism of λphage. The genetic elements used include a regulatory protein gene (merR) along with operator/promoter (O/P) derived from the mercury resistance operon from pDU1358 plasmid of Serratia marcescens. The expression of reporter gene gfp is also controlled by merR/O/P. Integration of the construct into the chromosome was done to increase the stability and precision of the biosensor. This biosensor could detect Hg(II) ions in the concentration range of 100-1700 nmol/L, and manifest the result as the expression of GFP. The GFP expression was significantly different (P ≤ 0.05) for each concentration of inducing Hg(II) ions in the detection range, which reduces the chances of misinterpretation of results. A model using regression method was also derived for the quantification of the concentration of Hg(II) in water samples.     

基于平面波导型荧光免疫传感器的双酚A检测适用性研究

双酚A是一种环境内分泌干扰物,逐渐受到国内外广泛关注.平面波导型荧光免疫传感器可以对水样中存在的痕量双酚A进行快速高灵敏度检测.在最优检测条件下测得双酚A标准曲线的检出限为(0.04±0.007)μg·L-1,线性区间为0.16~22.40μg·L-1,半抑制浓度为(1.67±0.47)μg·L-1.加入0.5%的EDTA到样品溶液中可以削弱水体硬度的干扰,并在最优条件下测得4种实际水样的加标回收率在88%~111%之间,相对标准偏差小于15%,表明该方法可以运用于实际水样中双酚A的检测.     

基于固定化AChE的流动注射型酶传感器研究

以固定化鲅鱼脑组织乙酰胆碱酯酶[acetylcholinesterase,AChE(EC 3.1.1.7)]为识别元件,以pH电极为换能器,构建流动注射型AChE酶传感器.该传感器在以磷酸盐缓冲液为载液的条件下具有良好的重现性(RSD=1.427%,n=10)和敏感性,可实现对有机磷化合物的在线监测;对甲基对硫磷具有线性响应的浓度范围为4.29×10-10^-4.29×10^-8mol·L^-1,最低检测限为1.3×10^-