萘降解质粒pND6-1中萘趋化基因nahY的克隆和核苷酸序列分析

假单胞菌（Pseudomonas）ND6菌株的萘降解基因位于102kb的质粒pND6-1上。以pUC18质粒为载体，制备了含有1.5-3.0kbpND6-1DNA随机片段的基因文库。通过DNA测序和DNA序列的同源性分析，从基因文库中筛选出含有萘趋化基因nahY的克隆。NahY基因的大小为1617bp，编码的萘趋化蛋白由538个氨基酸组成，与假单胞菌G7菌株的nahY基因相比，核苷酸序列的同源性是96.7％，编码的氨基酸序列的同源性是95％。图4参7     

华东地区某饮用水源地中磺胺类抗性基因的分布特征

饮用水源中检测到的抗生素抗性基因(antibiotic resistance genes,ARGs)对饮用水质安全和人体健康产生的潜在威胁受到广泛关注.在掌握了华东地区某饮用水源地13种磺胺类抗生素的污染特征基础上,进一步采用定性PCR和荧光定量PCR解析该饮用水源水和底泥中磺胺类ARGs(sul1、sul2)以及抗性基因可转移元件Ⅰ型整合酶基因(int I1)的分布特征.结果表明,3种基因在该饮用水源水和底泥中均100%检出,sul1基因是该饮用水源地中检出含量最高的磺胺类ARGs,在水源水中含量范围为1.5×104~6.4×105copies·mL~(-1),底泥中则高达1.6×108copies·g~(-1),较sul2、int I1基因分别高0.6~2.2、0.5~1.9个数量级.sul1、sul2和int I1基因在该水源地入水口和出水口处的绝对含量无显著差别,而在底泥中sul1、sul2和int I1基因的绝对含量则是出水口高于入水口.sul1在夏季水源地出水口的检出含量最高,为6.4×105copies·mL~(-1);int I1基因在冬季的检出含量高于其他季节.sul1基因与13种磺胺类抗生素具有相关性(r=0.69,P0.05),其中与磺胺甲唑的含量显著相关(r=0.79,P0.01);int I1与sul1、sul2的相对含量之间也存在正相关关系(r为0.80和0.73,P0.05),这表明int I1在磺胺类ARGs的水平转移过程中起到了重要作用.本研究为典型饮用水源地中ARGs的污染现状提供基础数据,也为管控饮用水环境的抗性基因污染和制定管理决策提供依据.     

Chromosomal genes conferring tolerance to heavy metal (Ag) toxicity

Bacterial strains were isolated from sediment samples from the Thames River. Successive transfer growth of the various strains on nutrient agar containing increasing concentrations of AgNO₃ revealed that three of the bacterial isolates were found to be capable of tolerating high concentrations of AgNO₃ ranging from 20 to 80 mM on a solid medium and up to 10 mM AgNO₃ in liquid medium. Molecular characterization and identification based on 16S rDNA gene sequencing of three strains of bacteria that are tolerant to silver nitrate showed that the major tolerant strains include the superbug, Shewanella oneidensis, Pseudomonas sp. and Bacillus sp. Protein extraction and two-dimensional (2D) sodium dodecyl sulfate SDS-polyacrylamide gel electrophoresis (PAGE) of the protein extracts in bacteria exposed to very high concentrations of AgNO₃ revealed a general reduction in the number of expressed proteins, although two protein spots were conspicuously over expressed in the exposed bacteria compared to control. The N-terminal amino acid sequence analysis of the protein spots identified the major up-regulated proteins as the outer membrane protein To1C (45.2 kDa) and the structural protein of the flagellar filament, flagellin (28.34 kDa), encoded for by the to1C and fliC genes, respectively. The roles of these genes in a number of multi-drug resistant pathogen and potentials for biotechnological applications in toxic metal control for treatment of contaminated ecosystems and biomining were discussed.     

好氧反硝化菌种DF2的分离鉴定及生理生化特性分析

分离了新型的高效反硝化细菌并研究其生物脱氮效率。通过观察形态学、生理生化鉴定和16SrRNA基因同源分析来鉴定菌种,致力于研究微生物污水脱氮处理的实际效果和克服反硝化过程中有害中间产物的积累。利用BTB培养基从水平潜流人工湿地基质中初步筛选出了9株平板阳性菌,革兰阴阳性均有。经试验研究发现该9种菌株在好氧条件下均具有一定的脱氮能力,其中以DF2和DF3 2种菌株脱氮能力最为显著,在3 d时间内NOX--N去除率达到了95%以上,NO2--N仅有微量积累;而其它7种菌株对NO3--N的去除率可以达到98%以上,但是NO2--N积累比较严重,积累量达到了NO3--N去除的50%~70%左右。经测序分析鉴定DF2和DF3分别属于德克斯氏菌属(Derxia)和假单胞菌属(Pseudomonadaceae)。德克斯氏菌属作为除氮的反硝化菌属在以往的研究中鲜有报道,该菌在好氧条件下可以合成周质NAR的亚基基因(napA)。     

硫自养反硝化颗粒表面与间隙微生物群落特征和基因分布

研究了单质硫颗粒自养反硝化柱中表面和间隙生物膜的微生物群落结构、功能基因和代谢途径等生物信息学特征.结果表明,硫颗粒表面生物膜的微生物菌群多样性低于间隙生物膜.氮代谢功能基因丰度差异较为显著,间隙生物膜中硝酸盐和亚硝酸盐的胞外转运蛋白基因丰度远高于表面生物膜,分别为0.0792%、0.109%与0.0157%、0.0314%.对于还原性反硝化代谢,表面生物膜的总基因丰度却明显低于间隙生物膜,分别为0.367%、0.406%.此外,参与反硝化过程的基因丰度明显不同,特别是将NO₃^-还原成NO₂^-以及将N₂O还原成N₂过程中的基因.对于硫代谢,没有观察到明显的差异.APS (硫酸腺苷)氧化是将SO₃^2-氧化为SO₄^2-的主要途径,其基因丰度远远高于直接氧化途径,分别为0.137%与0.0005%(表面)、0.138%与0.0007%(间隙).结果表明,在单质硫自养反硝化过程中,间隙生物膜与表面生物膜中的微生物存在合作关系,协同促进硫自养反硝化脱氮过程.     

Gene expression profiling of human liver carcinoma (HepG2) cells exposed to the marine toxin okadaic acid

The marine toxin, okadaic acid (OA) is produced by dinoflagellates of the genera Prorocentrum and Dinophysis and is the causative agent of the syndrome known as diarrheic shellfish poisoning. In addition, OA acts as both a tumor promoter, attributed to OA-induced inhibition of protein phosphatases as well as an inducer of apoptosis. To better understand the potentially divergent toxicological profile of OA, the concentration-dependent cytotoxicity and alterations in gene expression on the human liver tumor cell line HepG2 upon OA exposure were determined using RNA microarrays, DNA fragmentation, and cell proliferation assays as well as determinations of cell detachment and cell death in different concentrations of OA. mRNA expression was quantified for approximately 15,000 genes. Cell attachment and proliferation were both negatively correlated with OA concentration. Detached cells displayed necrotic DNA signatures but apoptosis also was broadly observed. Data suggest that OA has a concentration dependent effect on cell cycle, which might explain the divergent effects that at low concentration OA stimulates genes involved in the cell cycle and at high concentrations it stimulates apoptosis.     

上市航空公司盈利能力综合评价

自2008年以来,我国航空业一直处于发展低谷,评价航空公司的盈利能力及分析提高盈利能力的对策是一关键问题.许多学者习惯于应用单一的评价方法对航空上市企业进行评价,然而在对同一批航空上市企业进行评价时,不同的方法得出了不同的评价结果.基于此状况,提出一种系统的综合评价方法,并对全国12家典型航空上市企业进行综合评价,克服了以往应用单一方法在评价中的片面性,使评价结果更加全面客观.     

聚类GO术语在基因表达差异研究中的应用

针对基因功能分类体系基因本体(Gene Ontology,GO)特殊的有向无环图特点,改进传统的用单个GO术语检测基因差异表达信号的缺陷,设计出"聚类GO术语提升差异表达检测(ScaGO)"算法.通过简单的输入对照和实验组表达谱上的全部基因表达信号,来研究一些比较新的差异表达功能组,有助于进一步解释基因差异表达的生物学意义,如疾病发病机制、药物作用机理等.将ScaGO和基于单GO术语差异分析法应用到急性淋巴细胞性白血病数据集和酵母Rap1 DNA绑定突变体差异表达数据集上,结果显示,ScaGO能比基于单GO术语差异分析法发现一些新的与差异表达相关联的功能类基因,对于指导实验具有积极意义.图1表3参21