重组藻蓝蛋白与水稻类囊体膜之间光能传递的初步研究

为了研究藻蓝蛋白和高等植物类囊体膜之间的光能传递,本实验将诱导表达的重组藻蓝蛋白和水稻类囊体混合孵育后,检测其77K荧光发射光谱以及类囊体膜的电子传递速率和光化学量子产量,结果表明重组藻蓝蛋白和水稻类囊体膜光系统Ⅰ之间存在能量传递使得光化学量子产量提高.本研究为利用藻胆蛋白构建新型光合系统提供了理论基础和实验依据.     

Do endocrine disrupting chemicals threaten Mediterranean swordfish? Preliminary results of vitellogenin and Zona radiata proteins in Xiphias gladius

Endocrine Disrupting Chemicals (EDCs) have the potential to alter hormone pathways that regulate reproductive processes in wildlife and fishes. In this research the hypothesis that Mediterranean top predator species (such as large pelagic fish) are potentially at risk due to EDCs is investigated. These marine organisms tend to accumulate high concentrations of EDCs such as polyhalogenated aromatic hydrocarbons (PHAHs). The potential effects of EDCs on a fish species of commercial interest, the top predator Xiphias gladius (swordfish), were investigated using vitellogenin (Vtg) and Zona radiata proteins (Zrp) as diagnostic and prognostic biomarkers. Dramatic induction of typically female proteins (Vtg and Zrp) was detected by ELISA and Western Blot in adult males of the species. These results are the first warning of the potential risk for reproductive function of Mediterranean top predators, and suggest the need for continuous monitoring of this fragile marine environment.     

The role and significance of extracellular polymers in activated sludge. Part I: Literature review

In biological wastewater treatment, bacteria tend to aggregate by forming flocs, biofilms and even granules. A large part of the floc structure in activated sludge is composed of extracellular polymers. The extracellular polymeric substances (EPS) are mainly responsible for the structural and functional integrity of biofilms/flocs and they are considered to be important for the physico‐chemical properties of activated sludge. This review considers the composition of EPS and proportion between the constituents. Moreover it includes the relationship between the EPS constituents and sludge properties such as sludge retention time, hydrophobicity and surface charge.     

Seasonal Changes in Quality and Quantity of Food Available for Benthic Suspension-feeders in the Golfo Marconi (North-western Mediterranean)

Suspended particulate matter was collected, from the water layer at 10 cm above the sediments, over a period of 13 months in the Golfo Marconi (Ligurian Sea, NW Mediterranean). Measurements of seston concentration as well as the elemental (particulate organic carbon and nitrogen; POC and PON, respectively) and biochemical composition (lipids, proteins, carbohydrates, DNA) of particulate organic matter were carried out to assess quality and quantity of food potentially available to benthic suspension-feeders. Particulate organic matter showed wide qualitative and quantitative variations during the sampling year. Seston concentrations and POC did not reflect the quantity and quality of the food available to benthic suspension-feeders. The biopolymeric fraction of particulate matter (C-BPF, i.e. the sum of lipid, protein and carbohydrate carbon) was mostly composed of phytoplankton (which accounted for about 60% of C-BPF). The ratio of C-BPF to POC was utilized as a measure of the fraction which had the potential to be more readily available to consumers. Suspended organic matter showed higher values of the C-BPF:POC ratio during spring, and lower values in summer and autumn–winter. Quantitative estimates of the energy content of the suspended particulate matter were obtained from its biochemical composition. Bacterial dynamics were significantly related to changes in phytoplankton biomass. Bacteria accounted for a significant fraction of the biopolymeric carbon pool (annual average about 15%) and of the total particulate DNA (21·5%), thus enhancing the nutritional value of the particulate organic matter. The results achieved in this study indicate that the biochemical composition of the particulate matter provides additional information on the origin, quality and characteristics of the seston more readily available to benthic suspension-feeders.     

牙鲆芳香化酶Cyp19a多克隆抗体制备及其应用

芳香化酶Cyp19a在鱼类性别决定和性别分化中起关键作用,通过调控体内雌、雄激素的转化来影响鱼类性别表型形成。为深入研究Cyp19a在牙鲆(Paralichthys olivaceus)性腺分化和发育过程中的表达规律及作用机制,本文从牙鲆cDNA中克隆获得1557bp的cyp19a基因编码序列,并成功构建了原核重组表达质粒。经体外重组与纯化获得较高纯度的重组Cyp19a蛋白;以此作为抗原免疫兔子,制备多克隆抗体。用间接酶联免疫吸附剂测定(ELISA)技术检测抗血清效价,评估其免疫原性。结果表明免疫结束后得到的抗体血清效价超过了1∶50000,且纯化后抗体具有较好活性。Western Blot(WB)检测结果表明Cyp19a兔多抗可以特异性识别牙鲆重组Cyp19a蛋白和内源性Cyp19a蛋白。蛋白水平的雌雄性腺差异表达分析显示,牙鲆Cyp19a蛋白在卵巢中高表达,在精巢中微量表达。利用Foxl2和Dmrt1重组蛋白处理牙鲆性腺分化期幼鱼可分别显著上调和下调Cyp19a的表达(P<0.05)。综上所述,本研究成功制备了牙鲆Cyp19a多克隆抗体并进行了应用,为深入研究牙鲆等鱼类性别分化机制提供有力工具。     

刺参高温定向选育群体子一代耐温性状的分析

在室内控温条件下测定了定向选育获得的刺参(Apostichopus japonicus)耐高温群体子一代高温胁迫下的存活率及热休克蛋白(Hsp90和Hsp26)基因表达特征,从生理及分子水平分析了刺参耐高温群体子一代对高温胁迫的响应特征,确定高温耐受性的可遗传性。结果表明:与野生刺参群体子一代相比,定向选育刺参群体子一代在高温胁迫下表现出较高的存活率,32℃高温刺激下定向选育群体和野生群体稚参的存活率分别为80%和56.67%。高温胁迫下两个群体子一代的热休克蛋白90及26基因表达特征也存在差异,定向选育刺参群体子一代在高温刺激条件下热休克蛋白基因表达水平低于野生群体,且表达出现滞后现象。结果表明定向选育刺参群体子一代对高温具有一定的耐受能力,一定程度上验证了高温耐受性的可遗传性,该实验所获得的结果可以为后续刺参良种培育提供一定的理论依据和技术支撑。     

贻贝粘性蛋白组成、作用机理与应用

从贻贝足丝的结构和组成特征开始,全面总结了到目前为止所发现的贻贝足丝内部和表面层的结构特点以及相关重要组成蛋白的种类和理化特性,着重讨论了在贻贝足丝粘附中起重要作用的足丝斑块部分的粘性蛋白组成、结构、分布特征以及相互作用。同时,也讨论了贻贝粘附机理以及DOPA(3,4-二羟基苯丙氨酸)在实践中的重要应用,探讨了DOPA的作用及其衍生物在粘胶剂、密封剂、涂料、生物防污等领域,特别是生物医学中的应用和前景。     

三疣梭子蟹(Portunus trituberculatus)半乳糖凝集素PtGAL的基因克隆与原核重组表达

半乳糖凝集素(Galectin)是动物凝集素家族的一员,在脊椎动物和无脊椎动物中都参与了重要的免疫防御反应,是一种重要的免疫因子。通过RT-PCR和RACE技术克隆获得了三疣梭子蟹(Portunus trituberculatus)半乳糖凝集素基因全长,并命名为PtGal。该基因全长875bp,开放阅读框为669bp,编码222个氨基酸,包含一个糖识别结构域CRD(位于16—142位氨基酸),预测蛋白的分子量(Mw)为23529.4Da,理论等电点(p I)为5.21。同源性比对结果显示,PtGal编码的氨基酸序列与中华绒螯蟹、南美白对虾、日本囊对虾galectin的同源性分别为77%、66%、58%。实时荧光定量PCR(qRT-RCR)结果表明,PtGal在三疣梭子蟹肝胰腺、肌肉、肠、胃、鳃、心脏、性腺中都有表达,其中在性腺中的表达量最高,在肝胰腺中的表达量最低,但人工感染溶藻弧菌(Vibrio alginolyticus)后,肝胰腺的表达呈先升高后降低的趋势。利用原核表达系统对三疣梭子蟹PtGal进行了重组表达,并通过纯化复性获得了重组目的蛋白rPtGal,ELISA检测rPtGal与不同细菌及真菌的结合活性,结果表明,该重组蛋白与革兰氏阳性菌、阴性菌和真菌均有较强的结合。本研究为进一步深入研究三疣梭子蟹半乳糖凝集素的功能奠定了基础。