利用酵母双杂交系统筛选草鱼呼肠孤病毒NS38相互作用蛋白   总被引：2，自引：0，他引：2  

李杰  闫秀英  丁燏  吴灶和  简纪常  谢吉国 《海洋与湖沼》2013,44(2):305-309

采用酵母双杂交技术,对草鱼呼肠孤病毒096(GCRV096)非结构蛋白NS38相互作用的蛋白进行了筛选,并对阳性克隆中的插入序列进行测序与生物信息学分析。结果表明,含有重组质粒pGBKT7-NS38的酵母菌Y2HGold与含有草鱼肾脏细胞(CIK)cDNA文库质粒的酵母菌Y187融合成功,并筛选得到3株阳性克隆;发现两条不同的插入序列,其中一条序列编码的蛋白与40S核糖体蛋白S16亚基的同源性高达99%,而另一序列相应的蛋白不存在匹配的蛋白。该结果为进一步深入研究NS38蛋白在GCRV096复制过程中的生物学功能奠定了基础。  相似文献   

中国对虾肌动球蛋白变性后ATPase活性的研究   总被引：1，自引：0，他引：1  

林洪  王长峰  李兆杰  Ｋｈａｌｉｄ　 Ｊａｍｉｌ 《中国海洋大学学报(自然科学版)》1996,(4)

在中国对虾肌动球蛋白（Ａｏｔｏｍｙｏｓｉｎ）经冷冻变性和热变性后，对Ｃａ２＋－ＡＴＰａｓｅ、Ｍｇ２＋－ＡＴＰａｓｅ活性及蛋白质溶解度进行了研究。冷冻变性之后，肌动球蛋白Ｃａ２＋－ＡＴＰａｓｅ活性呈逐渐下降趋势，一般贮藏温度越低，活性降低越快。热变性之后，肌动球蛋白Ｃａ２＋或Ｍｇ２＋－ＡＴＰａｓｅ活性都随温度升高而下降，但肌动球蛋白Ｃａ２＋或Ｍｇ２＋－ＡＴＰａｓｅ活性在３５℃有一急剧下降的拐点，并分别在４５℃、４０℃附近低到０。由于蛋白质的变性，其溶解度也降低  相似文献   

养殖密度对循环水系统中大菱鲆生长和蛋白质代谢的影响     

韩岑  雷霁霖  刘宝良  贾睿  姜志强  赵奎峰  王国文 《海洋科学》2017,41(3):32-40

在工厂化循环水养殖系统中,将初始体质量为186 g±2.0 g的大菱鲆(Scophthalmus maximus)放养于低(9.4 kg/m~2±0.2 kg/m~2)、中(13.6 kg/m~2±0.8 kg/m~2)、高(19.1 kg/m~2±1.3 kg/m~2)3个养殖密度,以研究不同养殖密度对大菱鲆生长、消化酶和蛋白质代谢的影响。养殖120 d后,低、中、高试验组养殖密度分别增长至26.1 kg/m~2±1.2 kg/m~2、38.2 kg/m~2±2.5 kg/m~2、52.3 kg/m~2±3.6 kg/m~2。结果表明:低密度组和中密度组中大菱鲆增质量率、特定生长率、肥满度和蛋白质效率均显著(P0.05)高于高密度组;而饲料系数显著低于高密度组(P0.05)。低密度组和中密度组大菱鲆总蛋白酶和淀粉酶活力显著低于高密度组(P0.05);但脂肪酶活力在3个密度组之间无显著性差异。与低密度组相比,高密度组显著提高了谷氨酸脱氢酶活力,同时降低了谷草转氨酶和亮氨酸氨肽酶活力(P0.05),而对谷丙转氨酶活力无任何影响。综上所述,在工厂化循环水系统中,增加养殖密度能提高养殖的产量,但过高的养殖密度会对大菱鲆生长、消化酶活力以及蛋白质代谢产生不利的影响。  相似文献   

Effect of high salinity on cell growth and protein production of Antarctic ice microalgae Chlamydomonas sp. ICE-L     

郑洲  缪锦来  阚光峰  金青  丁燏  刘芳明  王守强  王以斌 《极地研究(英文版)》2010,21(1):81-90

Antarctic ice microalgae Chlamydomonas sp. ICE-L can survive and thrive in Antarctic sea ice. In this study, Chlamydomonas sp. ICE-L could survive at the salinity of 132‰ NaCl. SDS-PAGE showed that the density of 2 bands (26 and 36 kD) decreased obviously at the salinity of 99‰ NaCl compared to at the salinity of 33‰ NaCl. The soluble proteins in Chlamydomonas sp. ICE-L grown under salinity of 33‰ and 99% NaCl were compared by 2-D gel electrophoresis. After shocking with high salinity, 8 protein spots were found to disappear, and the density of 28 protein spots decreased. In addition, 19 protein spots were enhanced or induced, including one new peptide (51 kD). The changes of proteins might be correlated with the resistance for Chlamydomonas sp. ICE-L to high salinity.  相似文献   

Changes of proteins in the Antarctic ice microalga Chlamydomonas sp.cultured under UV-B radiation stress     

KAN Guangfeng  MIAO Jinlai  SHI Cuijuan  LI Guangyou 《海洋学报(英文版)》2006,25(2):135-141

1 IntroductionAntarctic ice m icroalga Chlamydomonas sp., akind ofgreen algae in the Antarctic sea ice, playsavery important role in Antarctic ecological environm ents.Antarctic“ozonehole”enhancesthesolarUVradiationarrivingatseaiceandcontinent. M anyphy…  相似文献   

太平洋牡蛎幼体附着变态差异蛋白质组研究          下载免费PDF全文

杨丙晔  濮菲  陈仲巍 《应用海洋学学报》2018,37(3):372-379

太平洋牡蛎(Crassostrea angulata)是中国主要的海洋养殖经济物种之一,具有重要的经济价值和商业价值,附着变态是牡蛎幼体从受精卵发育到成体所需的重要阶段,其分子机制的研究对牡蛎养殖育苗具有重要的指导意义.本研究利用双向电泳技术分离筛选牡蛎幼体附着变态过程中的差异表达的蛋白,进一步利用质谱技术鉴定幼体附着变态过程中发育相关的关键蛋白,共筛选出128个差异蛋白,鉴定并注释了39种蛋白.通过对差异表达蛋白功能分析,筛选获得了与纤毛组建相关的筑丝蛋白和与稳定钙离子胞内浓度相关的钙网蛋白,为揭示牡蛎幼体面盘组织及纤毛的退化消失和钙离子有效诱导幼体附着变态的机制提供分子数据.同时还获得了调节脂肪酸β-氧化、三羧酸循环和糖酵解重要生理功能的关键酶蛋白:乙酰辅酶A脱氢酶、异柠檬酸脱氢酶和3-磷酸甘油醛脱氢酶,以及调节无脊椎动物机体能量存储和释放的酶蛋白:精氨酸激酶和ATP合成酶,进一步阐明了牡蛎幼体在附着变态过程中能量调节的分子机制.  相似文献   

纳米银对青岛近海浮游细菌活性的影响研究     

何羿霖  白洁  白晓岩  李岿然 《中国海洋大学学报(自然科学版)》2020,50(4):76-82

纳米银广泛用于抗菌材料、医疗设备和其他抗菌产品。在纳米银产品的生产、运输、消费和处置过程中,纳米银将不可避免地释放到自然环境中,而水生生态系统是纳米银在自然界中重要的汇之一。本文以青岛近海为研究对象,采集现场海水进行实验室模拟培养,通过测定(48 h)不同浓度纳米银(0.5、5和50 mg/L)短期暴露下对表层海水中浮游细菌死亡率、蛋白质含量、碱性磷酸酶活性及氨肽酶活性的变化,研究纳米银对近岸海区浮游细菌生长和活性的影响。研究结果表明,在纳米银胁迫下细菌生长和活性受到显著抑制(P<0.05),高浓度组在培养4 h后死亡率达到最高值88.20%,随着暴露时间延长,纳米银胁迫组活细菌数量缓慢恢复、死亡率逐渐减小;纳米银对碱性磷酸酶活性和氨肽酶活性的抑制效应分别在培养4和8 h后达到最高,分别为8.58%和32.55%,在培养16 h后,酶活性开始缓慢上升,表现出对纳米银胁迫的适应性;纳米银组的蛋白质含量在培养8 h后达到最低,相比于对照组降低了29.80%,在培养后期同样有所升高,蛋白质含量接近于对照组。由此可见,纳米银对青岛近海浮游细菌的生长和活性都具有显著的抑制效应;不同浓度纳米银对细菌的抑制效应不同,纳米银浓度越高,抑制效应越明显(P<0.05),具有明显的浓度梯度效应;在培养后期,碱性磷酸酶活性和氨肽酶活性都有恢复,蛋白质含量上升,细菌死亡率下降,表明细菌对纳米银胁迫表现出一定的适应性。  相似文献   

Complete genome sequence and proteomic analysis of a thermophilic bacteriophage BV1     

LIU Bin  WU Suijie  XIE Lianhui 《海洋学报(英文版)》2010,29(3):84-89

Viruses of thermophiles are of great interest due to their roles in gene transfer, global geochemical cycle and evolution of life on earth. However, the thermophilic bacteriophages have not been studied extensively. In this investigation, a typical bacteriophage BV1 was obtained from a thermophilic bacterium Geobacillus sp. 6k512, which was isolated from an inshore hot spring in Xiamen of China. The BV1 contained a double-stranded linear DNA of 35 055 bp, which encodes 54 open reading frames (ORFs). Interestingly, eight of the 54 BV1 ORFs shared sequence similarities to genes from human disease-relevant bacteria. Seven proteins of the purified BV1 virions were identified by proteomic analysis. Determination of BV1 functional genomics would facilitate the better understanding of the mechanism for virus-thermophile interaction.  相似文献   

Three-dimensional fluorescence characteristics of dissolved organic matter produced by <Emphasis Type="Italic">Prorocentrum donghaiense</Emphasis> Lu     

赵卫红  王江涛  陈玫玫 《中国海洋湖沼学报》2009,27(3):564-569

Filtration and cross-flow ultrafiltration techniques were used to separate culture media of Prorocentrum donghaiense at the exponential growth,stationary and decline stages into <0.45 μm filtrate,100 kDa-0.45 μm,10-100 kDa and 1-10 kDa retentate and <1 kDa ultrafiltrate fractions.The fluorescence properties of different molecular weights of dissolved organic matter(DOM) were measured by excitation-emission matrix spectra.Protein-like and humic-like fluorophores were observed in the DOM produced by P.donghai...  相似文献   

Effect of lead on cytoskeletal protein stability in crucian carp Carassius auratus     

成嘉  张东裔  褚武英  刘芳  刘臻  周瑞雪  蒙涛  张建社 《中国海洋湖沼学报》2008,26(4)

Inorganic lead (Pb) is one of the most common environmental pollutants. Much evidence indicates that Pb exposure could directly affect fish growth and development. In this study, we investigated the cytotoxic effects of Pb on cytoskeletal protein stability at both protein and mRNA level in crucian carp Carassius auratus. Pb(NO3)2 treatment in concentration of 100 μmol/L resulted in decreased expression of both α- and β-tubulin but γ-tubulin as assayed with SDS-PAGE, Western Blot, and ELISA. In vivo and in vitro analyses on protein expression of tubulins are consistent. The effect of Pb on mRNA expression varied among different tissues. Our results suggest that cytotoxicity of Pb at protein translation level is stronger than at mRNA expression level.  相似文献