剑尾鱼卵黄蛋白原的ELISA检测

以卵黄脂磷蛋白(lipovitellin,Lv)抗血清为抗体,以纯化的卵黄蛋白原(vitellogenin,Vtg)为抗原,建立了间接酶联免疫吸附反应(ELISA)方法检测剑尾鱼(Xiphophorus helleri)雄鱼整体匀浆液中ρ(Vtg).结果表明,该方法的检测灵敏度为7.8 ng/mL,批内变异系数为5.094%,批间变异系数为5.540%,工作范围为32.5～2 000 ng/mL,在该范围内,标准曲线具有良好的线性和重复性.由于该方法可直接在1块或在不同的酶标板上准确地进行比较,因而可利用ELISA方法测定经诱导雄鱼整体匀浆液中ρ(Vtg)水平.结果显示,50 μg/L 17-β雌二醇(E₂)诱导21 d的雄性剑尾鱼有明显的Vtg产生;10 μg/L E₂诱导剑尾鱼亦有Vtg产生,但较50 μg/L E₂诱导组低;1 μg/L E₂诱导组及对照组剑尾鱼没有Vtg产生,检测孔OD450/对照OD₄₅₀(P/N)值小于2.1.      

人外周血淋巴细胞彗星试验检测饮用水水质毒性研究

应用人外周血淋巴细胞彗星试验对无锡充山水厂、中桥水厂,常州西石桥水厂进出水及金坛市拟用水源地长荡湖湖心水样的生物毒性进行了研究．结果表明,各水样的有机浓集物均可对人外周血淋巴细胞DNA产生不同程度的损伤,存在一定的遗传毒性．试验结果与各水样水质污染状况基本一致．同时,季节变化以及常规水处理过程对水样的生物毒性也有影响．研究表明,彗星试验可有效地用于检测饮用水质有机浓集物遗传毒性．     

Function of arsATorf7orf8 of Bacillus sp. CDB3 in arsenic resistance

Bacillus sp. CDB3 isolated from an arsenic contaminated cattledip site possesses an uncommon arsenic resistance (ars) operon bearing eight genes in the order of arsRYCDATorf7orf8. We investigated the functions of arsA, arsT, orf7 and orf8 in arsenic resistance using a plasmid-based gene knockout approach in the ars genedeficient Escherichia coli strain AW3110. The CDB3 arsA genewas shown to play a significant role in resistance, suggesting that the encoded ArsA may couplewith the arsenite transporter, forming an ArsAY complex that can enhance arsenite extrusion efficiency. Thedisruption of either arsT or orf7was not observed to affect arsenic resistance in the heterologous E. coli host, but their involvement in arsenic resistance can not be excluded. The orf8 gene is predicted to encode a putativedual-specificity protein phosphatasewhich also shares certain homology to arsenate reductases. The function loss of orf8 resulted in a remarkabledecrease in resistance to arsenate, though not arsenite. To examine if this effectwasdue to the reduction of arsenate by orf8, the arsC genewithin the 8-gene operonwasdisrupted. The resulting abolishment of arsenate resistance suggests that the involvement of orf8 in arsenic resistance is not via reductase activity.     

太湖不同湖区蓝藻细胞裂解速率的空间差异

2009年在太湖蓝藻水华形成初期(五月)、盛发期(九月)和衰亡期(十月和十一月),运用基于颗粒态酯酶,溶解性酯酶以及酯酶衰变常数测定的酯酶活性方法对不同湖区(藻型和草型湖区)蓝藻的细胞裂解速率进行了计算,在测定颗粒态酯酶、溶解性酯酶活性时,同步分析了太湖优势种群中蓝藻叶绿素a的含量.统计分析结果表明,叶绿素a的浓度与颗粒态酯酶、溶解性酯酶活性有很好的相关性,说明以酯酶活性为指标来计算太湖蓝藻细胞裂解速率是可行的.对不同湖区的细胞裂解速率进行比较,可见湖心和西太湖在蓝藻水华形成初期细胞裂解速率分别为0.072,0.048d-1.水华盛发期以及水华衰亡期,湖心和西太湖的细胞裂解速率分别为0.074～0.770d-1,0.014～0.110d-1.太湖湖心磷酸盐浓度比西太湖低,所以蓝藻生长速率慢,导致细胞裂解速率比西太湖高.但是,在梅梁湾和贡湖,衰亡末期磷酸盐浓度比其它月份高,细胞裂解速率也高.4个采样点在衰亡末期的细胞裂解速率比水华形成初期,暴发期和衰亡初期要高,可能的原因是气温和水体温度下降导致蓝藻生长速度减慢.本研究结果表明,太湖蓝藻细胞裂解速率有明显的空间差异,其具体的影响因素很多,营养盐只是其中一个.     

太湖梅梁湾有机污染物对纤细裸藻（Euglena gracilis）的遗传毒性效应

采用GC-MS方法分析了太湖梅梁湾水样的主要有机污染物成分和含量,共检测出15种目标化合物,表明该区域水体已经受到有机物污染;纤细裸藻(Euglena gracilis)在低浓度浓缩水样处理下(1倍组)生长无显著影响,叶绿素a、b含量和类胡萝卜素含量比对照组显著增加,而高浓度下(5倍和10倍组)生长明显受到抑制,叶绿素a和类胡萝卜素含量呈下降趋势;抗氧化酶系统中SOD和POD活性呈上升趋势,显示有机污染物胁迫可诱导抗氧化酶活性;彗星试验中,Olive尾矩和尾动量增加,纤细裸藻细胞DNA损伤程度随着太湖水样污染物浓度增加而加重,呈现出明显的剂量-效应关系,提示太湖梅梁湾水样具有潜在致突变性.结果表明,慧星试验和SOD活性试验结合使用在水环境中的遗传毒性监测方面具有较大的应用价值,适合作为水体有机污染监测的生物标志物.     

油田硫酸盐还原菌快速定量检测方法

现有油田废水中硫酸盐还原菌检测周期长、检测费用较高,本研究应用聚合酶链式反应(PCR)技术与倍比稀释法(MPN)相结合的DSR-MPN-PCR法,对硫酸盐还原菌进行快速定量检测.从废水中制备了直接用于PCR扩增的菌液,保证了定量准确性;建立以硫酸盐还原菌亚硫酸盐还原酶基因(Dsr)为靶位点的通用探针DSR1F和DSR5R的反应体系和扩增条件.结果表明,该方法检测灵敏度明显比液体稀释培养法高2个数量级,真实地表征了废水中实际的SRB菌数量,整个操作过程需要3～4　h,检测结果非常稳定,降低了检测费用,可以在生产中应用.     

两种兽药添加剂对蚯蚓的DNA损伤及其联合效应

采用土壤中的指示生物赤子爱胜蚓（Eisenia foetida）作为受试生物,研究了两种兽药添加剂四环素和金霉素对赤子爱胜蚓的急性毒性.结果表明：滤纸接触法染毒48h后,这两种兽药添加剂在所设最大受试剂量内没有引起蚯蚓死亡,半致死浓度LC50〉3.98×10^-2mg·cm^-2.应用彗星试验检测蚯蚓体腔细胞的DNA损...     

Assessment of source water contamination by estrogenic disrupting compounds in China

Detection of estrogenic disrupting compounds (EDCs) in drinking waters around China has led to rising concerns about health risks associated with these compounds. There is, however, a paucity of studies on the occurrence and identification of the main compounds responsible for this pollution in the source waters. To fill this void, we screened estrogenic activities of 23 source water samples from six main river systems in China, using a recombinant two-hybrid yeast assay. All sample extracts induced significant estrogenic activity, with E2 equivalents (EEQ) of raw water ranging from 0.08 to 2.40 ng/L. Additionally, 16 samples were selected for chemical analysis by gas chromatography-mass spectrometry. The EDCs of most concern, including estrone (E1), 17βup-estradiol (E2), 17αup-ethinylestradiol (EE2), estriol (E3), diethylstilbestrol (DES), estradiol valerate (EV), 4-t-octylphenol (4-t-OP), 4-nonylphenols (4-NP) and bisphenol A (BPA), were determined at concentrations of up to 2.98, 1.07, 2.67, 4.37, 2.52, 1.96, 89.52, 280.19 and 710.65 ng/L, respectively. Causality analysis, involving comparison of EEQ values from yeast assay and chemical analysis identified E2, EE2 and 4-NP as the main responsible compounds, accounting for the whole estrogenic activities (39.74% to 96.68%). The proposed approach using both chemical analysis and yeast assay could be used for the identification and evaluation of EDCs in source waters of China.     

Cloning,expression, and functional identification of CaSWEET7 and CaSWEET15 genes in Canarium album北大核心CSCD

糖外排转运蛋白(sugars will eventually be exported transporters,SWEET)介导植物光合同化产物蔗糖的跨膜运输.以橄榄(Canarium album(Lour.)Raeusch.)果实为材料,通过气相色谱串联质谱(GC-MS)检测橄榄果实中糖组分及含量变化,利用RT-PCR技术克隆得到CaSWEET7和CaSWEET15基因开放阅读框(ORF)序列.结果表明:CaSWEET7和CaSWEET15基因ORF全长分别为774 bp和951 bp,分别编码长度为257个和316个氨基酸残基,具有2个MtN3_slv结构域;进化树分析表明,CaSWEET7属于CladeⅡ,CaSWEET15属于CladeⅢ.实时荧光定量PCR(qRT-PCR)技术检测其在不同发育时期的果实中相对表达量变化,结果表明,随着果实发育,CaSWEET7和CaSWEET15表达量逐渐递增,且与果实发育蔗糖含量变化呈显著正相关(相关性系数分别为0.931和0.904,P<0.05);利用酵母功能互补证明CaSWEET7和CaSWEET15基因编码的蛋白具有转运蔗糖能力.本研究表明CaSWEET7和CaSWEET15基因可能在橄榄果实蔗糖积累中发挥作用.(图9表2参44)     

四膜虫彗星实验在环境水质检测中的应用

介绍四膜虫作为生物材料在彗星实验水质检测中的应用,探讨了将单细胞原生生物应用于彗星实验在环境水质检测、遗传毒性检测等方面的应用优势和发展前景。研究结果表明:过氧化氢浓度与细胞损伤之间呈现良好的剂量-效应关系,东湖水和长江水未造成四膜虫DNA的损伤,印染厂污水对细胞有严重的损伤作用,可能会影响正常的生态环境。