DNA polymorphism and globin chain analysis in the prenatal diagnosis of β-thalassaemia major in Taiwan

Thirty-six pregnancies in 25 families at risk of β-thalassaemia major received prenatal diagnosis. Chorionic villus sampling or amniocentesis was done in 35 pregnancies to obtain fetal cells for DNA linkage study, for which Southern blotting and DNA hybridization were used to detect seven restriction fragment length polymorphisms (RFLPs) within the β-globin gene cluster: ϵ-HincII, ^Gγ-HindIII, ^Aγ-HindIII, Φβ-HincII, 3′Φβ-HincII, β-AvaII, and 3′β BarnHI. β-Thalassaemia major was diagnosed in seven and excluded in 22 pregnancies. In the remaining six cases, β-thalassaemia major could not be excluded. In these six pregnancies and another one with late booking, ultrasound-guided cordocentesis was performed at the 22nd to 27th week of gestation. Globin chain composition was determined with urea-acetic acid-Triton X-100-12 per cent polyacrylamide gel electrophoresis. β-Thalassaemia major was diagnosed in two fetuses and excluded in the other five. Eleven fetuses (in which β-thalassaemia major was excluded) have been delivered and are healthy at more than 5 months old DNA linkage analysis coupled with globin chain electrophoresis provides an effective way for prenatal diagnosis of β-thalassaemia major, although these methods are being replaced by more direct detection techniques using oligonucleotide probes.     

单细胞凝胶电泳技术在环境污染物检测中的应用

主要介绍了单细胞凝胶电泳技术(SCGE),包括SCGE的原理、方法及其在环境污染物检测中的应用,该技术可用多种污染物的检测且灵敏度高,具有广阔的应用前景。即SCGE对大气污染物、重金属、醛类及辐射污染的检测,具有高度敏感性,是具有广阔应用前景的新技术。     

应用变性梯度凝胶电泳研究厦门西海域超微型真核浮游生物多样性

应用变性梯度凝胶电泳(DGGE)方法对厦门西海域超微型真核浮游生物的遗传多样性进行周年变化特征研究分析.结果表明,厦门西海域典型测站的超微型真核浮游生物群落具有丰富的多样性组成;厦门西海域超微型真核浮游生物群落遗传结构和主要类群组成季节性变化显著:不同典型测站之间的超微型真核浮游生物群落遗传结构的周年变化特征相似;不同典型测站之间的超微型真核浮游生物群落的类群组成也很相似,由绿藻、Stramenopiles、定鞭金藻、甲藻、未确定类群(unidentified groups)和其他类群组成.     

重金属Cr(Ⅵ)、Pb及Cu胁迫对双齿围沙蚕体腔细胞的DNA损伤

为探讨重金属Cr(Ⅵ)、Pb以及Cu对沙蚕体腔细胞DNA的毒性效应,以双齿围沙蚕为受试动物,重金属按不同剂量水平,Cr(Ⅵ):10、100和200 mg· L-1,Pb:5、50和100 mg·L-1,Cu:1、10和20 mg· L-1,分别胁迫沙蚕24 h,以不加任何重金属离子的海水为对照,采用单细胞凝胶电泳技术,检测其体腔细胞DNA损伤程度.结果表明,与空白对照组相比,3种重金属离子的各浓度组都能引起沙蚕体腔细胞DNA损伤,且3种重金属胁迫浓度与细胞DNA损伤程度之间存在显著的剂量-效应关系.双齿围沙蚕可以作为单细胞凝胶电泳的实验材料用于重金属所致环境污染的生物监测指示生物.     

两相一体式污泥浓缩消化(TISTD)反应器微生物多样性研究

针对课题组开发的两相一体式污泥浓缩消化反应器(TISTD),采用16S rDNA、PCR-TGGE等分子生物学技术,对反应器在不同阶段稳态下的内、外反应室中微生物的多样性和种群结构进行了研究.结果表明:TISTD反应器内外反应室中微生物群落均呈现高度的多样性分布,优势菌群明显,群落结构差异性很大,且外反应室较内反应室具有更多的微生物种群数量,外室是水解酸化的场所,污泥在外室经过微生物的水解酸化后进入内室,内室是严格厌氧的产甲烷场所;从反应器运行的不同时期来看,反应器中微生物优势种群呈现一定的差异,随着反应器负荷的改变,微生物的群落结构和种群数量也呈现出明显的演替过程,优势种群的功能地位处在动态变化中.微生物种群之间通过协同和竞争作用,形成了特定生态位的群落结构.研究结果解释了TISTD反应器同时浓缩和消化的工作机理,并为其推广、应用及优化调控提供了理论基础.     

Determination of vanillin in vanilla perfumes and air by capillary electrophoresis

The present study investigated capillary electrophoretic detection of vanillin in vanilla perfume and air. An UV-absorbance detector was used in a home-made capillary electrophoretic instrument. A fused silica capillary (outer diameter: 364μm, inner diameter: 50 m) was used as a separation capillary, and a high electric voltage (20 kV) was applied across the two ends of the capillary. Total length of the capillary was 70 cm, and the effective length was 55 cm. Experimental results showed that the vanillin peak was detected at about 600, 450, and 500 seconds when pH of running buffers in CE were 7.2, 9.3, and 11.5, respectively. The peak area of vanillin was proportional to its concentration in the range of 0-10^-2 mol/L. The detection limit was about 10^-5 mol/L. Vanillin concentration in a 1% vanilla perfume sample was determined to be about 3×10^-4 mol/L, agreed well with that obtained by a HPLC method. Furthermore, determination of vanillin in air by combination of CE and active carbon adsorption method was investigated.     

DNA提取方法对活性污泥微生物多样性PCR-DGGE检测的影响

评价超声波法、2种基于SDS的裂解法、改进的化学裂解法、微波法和冻融 玻璃珠 溶菌酶 SDS等6种DNA提取方法对活性污泥微生物多样性PCR-DGGE检测的影响.结果表明,2种不同的DNA纯化方式(溶液和胶纯化)及PCR扩增方式(直接和巢式PCR)对DGGE结果没有明显的影响,但不同的DNA提取方法对DGGE结果有明显的影响,其中2种基于SDS裂解法的DGGE条带较多,其提取的DNA产量(14.85μg/g湿重)和纯度(OD260:OD280＞1.80)也最高;改进的化学裂解法和微波法次之;超声波法、冻融 玻璃珠 溶菌酶 SDS法较少.研究表明,DNA提取方法是影响活性污泥微生物多样性PCR-DGGE检测结果的关键因素,基于SDS的裂解法是用于活性污泥微生物多样性PCR-DGGE研究的一种简便、可靠的总DNA提取方法.     

毛细管电泳中样品的在线富集

近年来,毛细管电泳以其速度快、效率高的分离分析技术受到越来越多领域的青睐,样品在线富集的方法有效的提高毛细管电泳由于进样体积小而造成的检测灵敏度难以提高的问题。文章简单介绍了毛细管电泳在线富集的原理,对其方法与应用进行了总结与概述。     

Effect of a high strength chemical industry wastewater on microbial community dynamics and mesophilic methane generation

A high strength chemical industry wastewater was assessed for its impact on anaerobic microbial com- munity dynamics and consequently mesophilic methane generation. Cumulative methane production was 251 mL/g total chemical oxygen demand removed at standard temperature and pressure at the end of 30 days experimental period with a highest recorded methane percentage of 80.6% of total biogas volume. Volatile fatty acids （VFAs） analysis revealed that acetic acid was the major intermediate VFAs produced with propionic acid accumulating over the experimental period. Quantitative analysis of microbial communities in the test and control groups with quantitative real time polymerase chain reaction highlighted that in the test group, Eubacteria （96.3%） was dominant in comparison with methanogens （3.7%）. The latter were dominated by Methanomicrobiales and Methanobacteriales while in test groups increased over the experimental period, reaching a maximum on day 30. Denaturing gradient gel electrophoresis profile was performed, targeting the 16S rRNA gene of Eubacteria and Archaea, with the DNA samples extracted at 3 different time points from the test groups. A phylogenetic tree was constructed for the sequences using the neighborhood joining method. The analysis revealed that the presence of organisms resembling Syntrophomonadaceae could have contributed to increased production of acetic and propionic acid intermediates while decrease of organisms resembling Pelotomaculum sp. could have most likely contributed to accumulation of propionic acid. This study suggested that the degradation of organic components within the high strength industrial wastewater is closely linked with the activity of certain niche microbial communities within eubacteria and methanogens.     

重金属Cr(VI)、Pb及Cu胁迫对双齿围沙蚕体腔细胞的DNA损伤

为探讨重金属Cr(VI)、Pb以及Cu对沙蚕体腔细胞DNA的毒性效应,以双齿围沙蚕为受试动物,重金属按不同剂量水平,Cr(VI):10、100和200 mg·L~(-1),Pb:5、50和100 mg·L~(-1),Cu:1、10和20 mg·L~(-1),分别胁迫沙蚕24 h,以不加任何重金属离子的海水为对照,采用单细胞凝胶电泳技术,检测其体腔细胞DNA损伤程度。结果表明,与空白对照组相比,3种重金属离子的各浓度组都能引起沙蚕体腔细胞DNA损伤,且3种重金属胁迫浓度与细胞DNA损伤程度之间存在显著的剂量-效应关系。双齿围沙蚕可以作为单细胞凝胶电泳的实验材料用于重金属所致环境污染的生物监测指示生物。