Detection of the end point temperature of thermal denatured protein in fish and chicken meat through SDS-PAGE electrophoresis

Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was applied in the detection of the end point temperature (EPT) of thermal denatured protein in fish and meat in this study. It was also used in studying the thermal denatured temperature range of proteins in salmon and chicken meat. The results show that the temperature ranges of denatured proteins were from 65 ℃ to 75 ℃ , and these temperature ranges were influenced by the processing methods. Through SDS-PAGE, the features of repeated he...     

6种海洋致病性弧菌36kDa外膜蛋白特性分析

研究本着筛选和制备弧菌共同外膜蛋白亚单位疫苗,用十二烷基肌氨酸钠抽提并结合超速离心的方法提取了鳗弧菌(Vibrio anguillarum)、鱼肠道弧菌(V.ichthyoenteri)、溶藻弧菌(V.alginolyticus)、副溶血弧菌(V.parahaemolyticus)、哈维氏弧菌(V.harveyi)和创伤弧菌(V.vulnificus)6种海洋致病性弧菌的主要外膜蛋白, SDS-PAGE图谱比较发现这6种弧菌都存在36 kDa的外膜蛋白条带.通过电泳洗脱分离纯化这6种弧菌的36 kDa外膜蛋白,并制备了鳗弧菌菌株SJ060621的36 kDa外膜蛋白多抗.ELISA和Western-blot印迹结果显示,仅鳗弧菌SJ060621、S010610-1和哈维氏弧菌的36 kDa外膜蛋白存在免疫交叉反应;双向电泳分析了鳗弧菌SJ060621、S010610-1、哈维氏弧菌和溶藻弧菌的36 kDa外膜蛋白,2-DE图谱显示出鳗弧菌SJ060621和S010610-1均由3种等电点相近的蛋白组成,存在很高的同源性,2株鳗弧菌与溶藻弧菌和哈维氏弧菌在蛋白质组成数量和等电点上都存在较大的差异,结果说明通过主要外膜蛋白的分子量和免疫特性研究,可为筛选不同弧菌共同外膜蛋白的亚单位疫苗提供有利参考.     

西施舌外套膜蛋白质组双向电泳体系的构建

建立了西施舌(Coelomactra antiquata)外套膜蛋白质组的双向凝胶电泳体系,比较分析了不同pH梯度胶条、水化条件的优化及不同染色方法对双向电泳结果的影响.结果表明:用pH 4～7的非线性胶条,水化时加上除盐步骤,采用硝酸银染色,即可获得高分辨率、高重现性的双向电泳图谱,为进一步研究西施舌外套膜蛋白质的组成奠定了基础.     

黄海西北近岸沉积物中细菌群落空间分布特征

采用16S rDNA文库和变性梯度凝胶电泳(DGGE)技术,对黄海西北部3个近岸站位沉积物中细菌群落多样性及空间分布特征进行了调查和解析。对表层沉积物16S rDNA序列统计表明,各站位细菌群落多样性很高,γ-和δ-变形菌纲分别占克隆序列总数的20%~32%,是沉积物中的绝对优势类群。DGGE图谱分析表明,同一站位中不同深度的细菌群落结构相似性较高,而不同站位间群落结构相差较远。研究表明在黄海西北近岸沉积物中细菌群落多样性较高,优势类群明显,在较小尺度范围内群落结构的垂直变化不明显。     

Similarity of electrophoretic dissolved protein spectra from coastal to pelagic seawaters

Dissolved proteins in seawater samples collected from a coastal area of Tokyo Bay, Sagami Bay and a location off the Kuroshio Current were investigated by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and high resolution two-dimensional electrophoresis (2-DE). Four to nine protein bands were detected in SDS-PAGE in the apparent molecular weight (MW) range from 12 kilo Dalton (kDa) to 49 kDa. The 2-DE technique distinguished 10 to 46 protein spots exhibiting isoelectric point (pI)/MW ranging 4.3–9.2/12–63 kDa. The elecrophoretic patterns were similar between the coastal and pelagic samples, as well as previously reported patterns from various pelagic areas. The close similarity of electrophoretic mobility on both SDS-PAGE and 2-DE gels indicates the compositional homogeneity of dissolved proteins in seawater throughout a broad range of marine environments. Proteinaceous dissolved organic matter (DOM) that was unresolved and smeared staining characteristics on both SDS-PAGE and 2-DE gels was first observed in Tokyo Bay waters in the present study and its possible sources are discussed. Although the two protein species, 48 kDa and 39 kDa proteins, have been identified as homologues of Porin P and low molecular weight-alkaline phosphatase of Pseudomonas aeruginosa PAO1, respectively, four strains of P. aeruginosa and two species of Pseudomonas spp. have been newly identified as the source organisms of these proteins using the N-terminal amino acid sequence data determined in previous studies.     

日本沼虾黑鳃病几种同工酶的变化与病理分析

酯酶（EST），过氧化物酶（POD），乳酸脱氢酶（LDH）和苹果酸脱氢酶（MDH）同工酶在日本沼虾中存在着组织和器官的特异性，患黑鳃病后酶带发生显著变化。在肌肉、肝脏、心脏和鳃中病虾较健康虾的LDH酶谱分别缺失了2、6、2和1条酶带，同时肝脏中新增了1条酶带；POD酶谱分别缺失了0、7、4和2酶带，肌肉中新增了2条酶带；EST酶谱分别缺失了0、2、3和1条酶带，肌肉、心脏和鳃中分别新增了1、2和2条酶带；MDH酶谱分别缺失了0、2、1和1条酶带，肌肉和脏脏中各新增了1条酶带。     

Spatial and temporal changes in microbial diversity of the Marmara Sea sediments

Spatial (10 different locations) and temporal (2 years) changes in characteristics of the Marmara Sea Sediments were monitored to determine interactions between the chemical and microbial diversity. The sediments were rich in terms of hydrocarbon, nitrate, Ni and microbial cell content. Denitrifying, sulfate reducing, fermentative and methanogenic organisms were co-abundant in 15 cm below the sea floor. The local variations in the sediments’ characteristics were more distinctive than the temporal ones. The sulfate and nitrate contents were the main drivers of the changes in the microbial community compositions. N and P were limited for microbial growth in the sediments, and their levels determined the total cell abundance and activity. Seasonal shifts in temperatures of the shallow sediments were also reflected in the active cell abundances. It was concluded that the Marmara Sea is a promising ecosystem for the further investigation of the ecologically important microbial processes.     

几种养殖鲍同工酶生化遗传的比较研究

运用聚丙烯酰胺凝胶电泳对我国主要养殖鲍(皱纹盘鲍、盘鲍及九孔鲍)12种同工酶(LDH、ME、MDH、GPI、PGM、IDH、IDDH、SOD、EST、ODH、ADH、AAT)26个基因座位进行了分析．结果表明：EST、PGM可以作为区分三种(亚种)鲍的生化遗传标记；在所分析的位点中，未发现多态现象，表明三种养殖鲍杂合子缺失严重，其主要原因可能在于哑基因、近交与自然选择。