利用乙酰胆碱酯酶传感器监测海水久效磷

用戊二醛(GA)将乙酰胆碱醇酶(ACHE)交联在透析膜上,并与pH电极结合,由此开发出一种基于固定化酶的电位型传感器。该酶电极可以快速、灵敏地测定海水中的久效磷。仪器对海水中10^-10～10^-7的久效磷农药具有极好的线性响应,检测下限为0．1μg／L,加标回收率在90％～103％之间,分析1个海水样品只需50min。     

BOD生物传感器在海洋监测中的应用

选用一株耐高渗透压的酵母菌作为敏感材料可以实现BOD的快速测定。采用聚乙烯醇包埋方法对酵母菌进行包埋 ,可在 15min内完成一个海水BOD标准样品测量 ,测定结果具有很好的线性关系。并研究了对高分子有机物的响应、使用时间及保存方式     

Runoff rates,chemical speciation and bioavailability of copper released from naturally patinated copper

The release of copper, induced by atmospheric corrosion, from naturally patinated copper of varying age (0 and 30 years) has been investigated together with its potential ecotoxic effect. Results were generated in an interdisciplinary research effort in which corrosion science and ecotoxicology aspects were combined. The aim of the investigation was to elucidate the situation when copper-containing rainwater leaves a roof in terms of runoff rate, chemical speciation, bioavailability and ecotoxicity effects. Data have been collected during a three-year field exposure conducted in the urban environment of Stockholm, Sweden. The potential environmental effects have been evaluated using a combination of a copper specific biosensor test with the bacterium Alcaligenes eutrophus and the conventional 72-h growth inhibition test with the green alga Raphidocelis subcapitata. The results show annual runoff rates between 1.0 and 1.5 g/m2 year for naturally patinated copper of varying age. The runoff rate increased slightly with patina age, which mainly is attributed to the enhanced first flush effect observed on thicker patina layers. The total copper concentration in investigated runoff samplings ranged from 0.9 to 9.7 mg/l. Both computer modeling and experimental studies revealed that the majority (60-100%) of released copper was present as the free hydrated cupric ion, Cu(H2O)6(2+), the most bioavailable copper species. However, other copper species in the runoff water, such as, e.g. Cu(OH)+ and Cu2(OH)2(2+), were also bioavailable. The copper-containing runoff water, sampled directly after release from the roof, caused significant reduction in growth rate of the green alga. It should be emphasized that the results describe the runoff situation immediately after release from the copper roof and not the real environmental ecotoxicity. Therefore the data should only be used as an initial assessment of the potential environmental effect of copper runoff from building applications. Future risk assessments should also consider dilution effects of copper, changes in its chemical speciation and bioavailability during environmental entry, and type and sensitivity of the receiving ecosystem.     

绿色荧光蛋白分子标记在环境微生物学研究中的应用

绿色荧光蛋白(green fluorescent protein,GFP)分子标记是现有遗传标记中最简单方便的一种方法，GFP及GFP突变体在微生物降解污染物、生物膜菌群构架、环境生态学和环境检测生物传感器等研究领域取得了很好的应用效果。     

Preimplantation genetic diagnostic protocols for α- and β-thalassaemias using multiplex fluorescent PCR

Haemoglobinopathies including α- and β-thalassaemia are the world's most common class of single gene disorder. Prenatal diagnosis (PND) for β-thalassaemia has been proven to be an effective strategy for controlling the incidence of new cases and is widely used in several countries where the disease is common. Successful preimplantation genetic diagnosis (PGD) protocols for β-thalassaemia have been introduced using restriction fragment length polymorphism (RFLP), single-stranded conformation polymorphism (SSCP) and denaturing gradient gel electrophoresis (DGGE). However, contamination and allele dropout (ADO) remain an important concern for all of these strategies. In the present study two PGD protocols for detecting β-thalassaemia mutations (codon 41-42 and IVSI-110) and one for α-thalassaemia (SEA mutation) have been designed and tested. These methods contain failsafe mechanisms to reduce the risk of misdiagnosis due to ADO or contamination and utilise multiplex fluorescent PCR (F-PCR). Interestingly, amplification efficiency and ADO were significantly affected by the choice of DNA polymerase and the freshness of the single cells used. The close similarity between the DNA sequences of β-globin and δ-globin was also found to be an important issue that necessitated careful design of primers for the β-globin gene. Copyright © 2001 John Wiley & Sons, Ltd.     

生物传感细胞ADP1_pWHlux在水环境急性毒性检测中的应用

针对天然发光菌和以模式微生物为宿主构建的生物传感细胞在急性毒性检测应用中对测试条件要求苛刻等适用性问题,本研究将1株基因工程构建的生物传感细胞不动杆菌ADP1_p WHlux应用于急性毒性检测,建立检测方法,考察其灵敏度及检测范围.结果表明,ADP1_p WHlux发光受急性毒物的抑制,毒物剂量与发光抑制存在剂量效应关系.在4 mg·L-1Hg Cl2诱导下仅5 min可作出响应,暴露30~60 min后可以给出较为准确的结果.对Hg Cl2的检出限可达0.04 mg·L-1.对我国生活饮用水卫生标准的指标中Be2+、Ba2+、Cu2+、Ni2+检出效果明显,对Be2+、Ba2+、Cu2+的检测范围均在0.025~250 mg·L-1,对Ni2+的检测范围在0.002 5~250 mg·L-1,对Pb2+、Br O-3、Cl O-2的检出限均在0.002 5 mg·L-1,对Cl O-3检出限为0.025mg·L-1.采用ADP1_p WHlux生物传感细胞检测方法对北京市清河水环境急性毒性进行评价,表明ADP1_p WHlux生物传感细胞检测方法可用于污染水样检测.     

铜陵市河流沉积物中硝化和反硝化微生物分布特征

矿区河流不仅受到矿业活动带来污染物的影响,还受到区域内生活污水及农田退水中氮、磷和有机物的污染.本研究以铜陵市河流表层沉积物为研究对象,采用荧光定量PCR技术分析季节和污染类型对沉积物样品中硝化(amo A)和反硝化(nir S和nir K)功能基因丰度的影响.结果表明,沉积物样品中氨氧化古菌(AOA)和细菌(AOB)基因丰度变化范围分别为1.74×105~1.45×108copies·g-1和1.39×105~3.39×107copies·g-1,AOA平均丰度是AOB的4.39倍;反硝化基因(nir S和nir K基因)丰度变化范围分别为1.69×107~8.55×109copies·g-1和4.45×106~1.51×108copies·g-1,nir S基因平均丰度为nir K基因的28.35倍.沉积物AOA丰度呈现春季和秋季较高,夏季和冬季较低的趋势,而AOB在春季和冬季沉积物中的丰度高于夏季和秋季;反硝化基因丰度则表现为春季(nir S)/秋季(nir K)夏季冬季秋季(nir S)/春季(nir K).矿区周围以重金属为主要污染物的河流沉积物中两种amo A和nir S基因的丰度高于毗邻农田区域主要受到氮、磷及有机物污染的河流,后者的nir K基因丰度更高.     

First application of preimplantation genetic diagnosis to neurofibromatosis type 2 (NF2)

Neurofibromatosis type 2 (NF2) is a dominantly inherited cancer predisposition syndrome that is caused bymutations in the NF2 gene. We report here the first clinical preimplantation genetic diagnosis (PGD) forNF2. A protocol was developed to simultaneously amplify the mutation and a single nucleotide polymorphism (SNP) located within the gene. The mutation and polymorphism were analysed by simultaneous fluorescent single-strand conformation polymorphism (SSCP) on an automated DNA sequencer. The mutation, carried by the male partner, was a single base pair substitution affecting a splice site in intron 4 of the gene. The female partner was infertile due to polycystic ovary syndrome and would require IVF to conceive. The couple was found to be informative at a linked intragenic SNP situated in the 5′ untranslated region of the gene. The SNP was included in the assay to reduce the risk of misdiagnosis due to allele dropout (ADO). The couple underwent three cycles of treatment during which a total of 43 blastomeres were biopsied from 31 embryos. Amplification at both loci was obtained in 35 cells (81%). A total of five embryos were transferred, two in the first cycle, two in the second and one in the third. No pregnancy ensued. The results of the diagnoses indicated that, in this couple, the inheritance of the mutation may be non-Mendelian. Out of a total of 32 embryos tested only four were found not to carry the mutation. The reasons for this apparent skew remain unknown. Copyright © 2002 John Wiley & Sons, Ltd.     

太湖草、藻湖区沉积物WSOM组成和分子结构分析

草型湖泊（区）和藻型湖泊（区）是浅水湖泊的两种典型状态,两种湖泊中有机质（OM）的来源和转化存在明显差异.有机质在沉积物上的埋藏是湖泊碳汇功能的重要体现.沉积物中的有机质可能发生后续的转化,影响湖泊碳汇功能;其转化过程与其固有的性质密切相关.目前为止,有关草型和藻型湖泊沉积物中有机质的组成,特别是分子结构差异的信息较少.为此,采集太湖草型湖区（东太湖）和藻型湖区（梅梁湾）沉积物,提取和纯化沉积物中的水溶性有机质（分别记为M-WSOM和A-WSOM）,并采用紫外-可见吸收光谱、三维荧光光谱、傅立叶红外光谱和傅里叶变换-离子回旋共振-质谱（FT-ICR MS）对WSOM性质组成和分子结构进行详尽的表征与分析.结果表明：①M-WSOM的相对分子质量比A-WSOM的（E₂ ：E₃指标）稍大;FT-ICR MS的数据显示,M-WSOM和A-WSOM的相对分子质量分别为388.9和379.9.②M-WSOM的芳香性略高于A-WSOM的（SUVA₂₅₄和HIX指标）;同样地,FT-ICR MS的结果显示,M-WSOM中稠环芳烃（6.3%,强度加权平均占比,下同）、芳香类物质（7.7%）的含量高于A-WSOM （分别为1.1%和4.4%）;③荧光组分结果表明A-WSOM中类蛋白质组分的含量大于M-WSOM;FT-ICR MS的结果与之对应,A-WSOM的多肽含量（35.5%）高于M-WSOM的（15.6%）;④质谱数据显示了高分辨的分子信息：M-WSOM和A-WSOM中,沉积物中含杂原子的分子分别占82.9%和91.7%;其中含氮元素的分子占比高达53.5%和78.5%;含磷元素分子占30.4%和41.4%,M-WSOM磷元素主要分布在脂肪烃和高度不饱和低氧物质中,而A-WSOM则主要分布在多肽中.揭示了浅水湖泊两种典型状态沉积物OM的组成和分子结构;有助于研究湖泊沉积物的温室气体排放,及碳、氮、磷元素的生物地球化学过程.