Methods Comparison for Microsatellite Marker Development： Different Isolation Methods, Different Yield Efficiency     

Aibin Zhan  Zhenmin Bao  Xiaoli Hu  Wei Lu  Jingjie Hu 《中国海洋大学学报(英文版)》2009,8(2):161-165

Microsatellite markers have become one kind of the most important molecular tools used in various researches. A large number of microsatellite markers are required for the whole genome survey in the fields of molecular ecology, quantitative genetics and genomics. Therefore, it is extremely necessary to select several versatile, low-cost, efficient and time- and labor-saving methods to develop a large panel of microsatellite markers. In this study, we used Zhikong scallop (Chlamys farreri) as the target species to compare the efficiency of the five methods derived from three strategies for microsatellite marker development. The results showed that the strategy of constructing small insert genomic DNA library resulted in poor efficiency, while the microsatellite-enriched strategy highly improved the isolation efficiency. Although the mining public database strategy is time- and cost-saving, it is difficult to obtain a large number of microsatellite markers, mainly due to the limited sequence data of non-model species deposited in public databases. Based on the results in this study, we recommend two methods, microsatellite-enriched library construction method and FIASCO-colony hybridization method, for large-scale microsatellite marker development. Both methods were derived from the mi-crosatellite-enriched strategy. The experimental results obtained from Zhikong scallop also provide the reference for microsatellite marker development in other species with large genomes.  相似文献   

Isolation and Characterization of Collagen from Squid(Ommastrephes bartrami) Skin   总被引：1，自引：0，他引：1  

YAN Mingyan  LI Bafang    ZHAO Xue College of Food Science  Engineering  Ocean University of China  Qingdao  P. R. China 《中国海洋大学学报(英文版)》2009,(2)

Collagen of squid(Ommastrephes bartrami) skin was examined in the present study. Histology showed that collagen fiber in the skin was partially cross-linked with muscle fiber. Acid-solubilized collagen(ASC) and pepsin-solubilized collagen(PSC) were extracted from the skin and characterized. The results of amino acid composition and electrophoretic patterns revealed that ASC and PSC were both type Ⅰ collagen,containing α1 and α2 chains. FTIR(fourier transform infrared spectroscopy) investigations confirmed t...  相似文献   

高效分解水产原料的菌株分离鉴定和组合发酵初探     

宋敏  刘为  张鹰炯  童晓倩  李鹏  王斌  罗红宇 《海洋与湖沼》2015,46(4):800-807

为从一种组成未知的商用复合菌中分离并筛选出优势发酵菌株,本实验采用稀释平板培养法,对菌种进行分离纯化,同时参照《真菌鉴定手册》和《常见细菌鉴定手册》,根据其形态学特征、生理生化特性,并分别结合16S r DNA和18S r DNA序列的比对分析对菌种进行鉴定。另外,本研究通过生长曲线测定,单菌种发酵试验以及多菌种混合发酵发酵试验,以游离氨基酸态氮为指标,以期筛选出发酵低值水产品制备菌肥的最佳菌株组合。结果显示,共分离得到4株菌,分别为a热带假丝酵母、b罗伦隐球菌、c枯草芽孢杆菌、d蜡样芽胞杆菌。且菌种组合acd发酵后游离氨基酸态氮的含量较对照组上升最显著,为8.276g/L。故可确定最佳发酵菌株配方为acd(热带假丝酵母+枯草芽孢杆菌+蜡样芽胞杆菌)。  相似文献   

Protracted oogenesis and annual reproductive periodicity in the deep‐sea pennatulacean Halipteris finmarchica (Anthozoa,Octocorallia)          下载免费PDF全文

Sandrine Baillon  Jean‐François Hamel  Annie Mercier 《Marine Ecology》2015,36(4):1364-1378

Halipteris finmarchica is one of the most common species of deep‐sea pennatulacean corals in the Northwest Atlantic; it was recently determined to act as a biogenic substrate for other species and as a nursery for fish larvae. Its reproductive cycle was investigated in colonies sampled in 2006 and 2007 along the continental slope of Newfoundland and Labrador (Canada). Halipteris finmarchica exhibits large oocytes (maximum diameter of 1000 μm), which are consistent with lecithotrophic larval development. Female potential fecundity based on mature oocytes just before spawning was ~6 oocytes · polyp^?1 (500–6300 oocytes · colony^?1); male potential fecundity was 16 spermatocysts · polyp^?1 (5500–57,400 spermatocysts · colony^?1). Based on statistical analysis of size‐probability frequency distributions, males harboured one cohort of spermatocysts that matured inside 8–11 months, whereas females harboured two distinct cohorts of oocytes; a persistent pool of small ones (≤400 μm) and a small number (~20%) of larger ones that grew from ~400 to >800 μm over a year. Despite this difference in the tempo of oogenesis and spermatogenesis, a synchronic annual spawning was detected. A latitudinal shift in the spawning period occurred from south (April in the Laurentian Channel) to north (May in Grand Banks and July–August in Labrador/Lower Arctic), following the development of the phytoplankton bloom (i.e. sinking of phytodetritus). Prolonged oogenesis with the simultaneous presence of different oocyte classes in a given polyp is likely not uncommon in deep‐sea octocorals and could hamper the detection of annual/seasonal reproduction when sample sizes are low and/or time series discontinued or brief.  相似文献   

一株拮抗菌C-5 的分离及鉴定   总被引：1，自引：1，他引：0  

韦露  陈偿  龙云映  蔡奕明 《海洋科学》2015,39(10):29-35

针对目前海参养殖中水产微生态制剂品种单一、施用效果不稳定等现状,作者从地瓜参(Acaudina molpadioidea)肠道分离土著肠杆菌(Enterobacter),筛选到1株具有较好拮抗作用的海洋细菌C-5,该菌对副溶血弧菌(Vibrio parahaemolyticus)、溶藻弧菌(Vibrio alginolyticus)、杀鲑气单胞菌(Aeromonas salmonicida)等7种细菌有拮抗作用;通过形态观察、生理生化和16S rDNA基因序列分析鉴定为霍肠杆菌(Enterobacter hormaechei);通过卤虫(Artemia)和药敏实验检测该菌具有安全性,其对卤虫的72 h半致死浓度(LC50)为108cfu/m L,只对20种常规抗生素中的5种产生耐药性;对C-5的发酵条件进行优化,该菌优化后的发酵条件为:温度30℃,pH 7.0,盐度2%。结果表明,C-5有望作为一种益生菌应用于海水养殖病害控制。  相似文献   

海洋红藻岩生多管藻绒毛变种Polysiphonia scopulorum var. villum形态学研究          下载免费PDF全文

丁兰平  钟丽敏  谭华强  黄冰心 《热带海洋学报》2015,34(1):42-48

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隔离变压器在电源系统防雷中的应用     

周方君  周萌  张佳晖  万瀚仁  汪胜宝 《气象科技》2015,43(5):969-972

分析了危害弱电设备的主要电涌类型及特点,探讨了在实际防雷工程中低压配电系统中常用的氧化锌（ZnO）电涌保护器（SPD）存在启动电压偏高,对耐过电压能力较低的电气电子设备存在对电涌防护不到位的问题;将隔离变压器作为防电涌器件引入到弱电设备防雷工程中,利用初级绕组与次级绕组之间只有电磁耦合,无直接电气通道的特性隔离共模过电压;利用铁芯的磁饱和、磁滞现象抑制和削减雷击感应过电压的差模分量。将隔离变压器用于氧化锌电涌保护器之后作为精保护单元,弥补氧化锌电涌保护器的缺点,使弱电设备得到可靠保护,经实际运用验证了它的防雷效果。  相似文献   

基于高通量测序的象山港水域细菌多样性的研究     

刘岩  李志恩  袁一鸣  张真  钱鹏宇  叶欢  刘松怡  王泽  韩姣姣  周君  苏秀榕 《海洋与湖沼》2022,53(3):645-653

象山港位于浙江省宁波市东南部沿海,属于半封闭式海湾,是以海水养殖区和海洋牧场示范区为主的重要多功能水域,近年来生态环境问题日益突出。为研究象山港海域在早春低水温环境中细菌的多样性,通过高通量测序方法对3月份的象山港水域进行检测。沿象山港南部海湾随机选取6个取样点(G04,G06,G08,G09,G011,G012),共检出836属和2 166种细菌,隶属33个门、91纲、186目和352科。其中优势细菌类群主要包括变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、放线菌门(Actinobacteria)、厚壁菌门(Firmicutes)和蓝细菌门(Cyanobacteria)。变形菌门在G04中占比最高(91.45%),拟杆菌门在G09中占比最高(18.05%);而放线菌门在G12中占比最高(7.95%)。在经度较高且离海岸较远的水域,细菌丰富度也越高;同时随着取样位点纬度的增加,γ-变形菌丰度逐步增高,其中在G04中最高(72.76%),在G12中最低(29.11%)。各取样点的细菌分离培养结果也表明弧菌属丰度较高,且主要集中在G04中。利用高通量测序方法揭示了象山港早春全水域的细菌结构组成框架图,可为象山港的水产养殖和可持续发展提供参考依据。  相似文献   

海洋来源羽毛降解菌株的筛选、鉴定及酶学性质研究          下载免费PDF全文

张红岩  何瑾  杨梦莹  张妮  王芳  申乃坤 《应用海洋学学报》2022,41(2):338-346

微生物降解羽毛既可保护环境又能实现废物的资源化利用,具有显著的经济效益和社会效益。本研究从广西钦州港海鸭养殖淤泥筛选获得一株可高效降解羽毛的菌株Gxun-20,通过对该菌株形态特征、生理生化特性和16S rRNA基因序列进行分析,以确定菌株的分类,并对菌株产角蛋白酶的条件及酶学性质进行研究。结果表明：该菌为副短短芽孢杆菌(Brevibacillus parabrevis);菌株最适产酶条件为：温度34 ℃,初始pH 6.5,羽毛含量1.5%(质量分数),发酵48 h,角蛋白酶活性可达277.45 U/mL。菌株Gxun-20所分泌的角蛋白酶最适温度为50 ℃,且热稳定性较好,最适pH为7.5;金属离子Fe³⁺、Cu²⁺、Co²⁺、Fe²⁺和Mn²⁺对酶活性有显著抑制作用,而Ca²⁺、Na⁺对酶活性有明显促进作用;金属螯合剂乙二胺四乙酸(ethylene diamine tetraacetic acid, EDTA)可显著抑制酶的活性,蛋白酶抑制剂苯甲基磺酰氟(pheny methyl sulfonyl fluoride, PMSF)可完全抑制酶活性,表明该蛋白酶的活性中心含有金属离子和丝氨酸;异丙醇可使酶活性提高至129.34%,酶活性在十二烷基硫酸钠(sodium dodecyl sulfate, SDS)、吐温40溶液中保持稳定;酶的底物特异性结果表明,该酶对羽毛类β-角蛋白及酪蛋白降解能力较好,而对牛血清蛋白、I型胶原及人发类α 角蛋白降解能力较弱。海洋来源的副短短芽孢杆菌Gxun-20及其所产角蛋白酶在废弃羽毛降解方面具有良好的应用价值。  相似文献   

太平洋鳕鱼脑中硫苷脂的分离纯化和分析     

薛长湖  宋雨  徐杰  贾子才  丛培旭 《中国海洋大学学报(自然科学版)》2014,44(10):51-56

以太平洋鳕鱼为原料,从鳕鱼脑中分离硫苷脂,确立了提取及纯化的条件,并对硫苷脂的纯度和分子种进行分析。首先,采用氯仿/甲醇(2∶1,v/v)提取总脂。然后,依次采用氯仿/甲醇/水(7∶3∶0.3,v/v/v)洗脱硅胶柱层析,含0.2mol/L乙酸铵的氯仿/甲醇/水(30∶60∶8,v/v/v)洗脱DEAE Sephadex-A25离子交换柱层析,40%甲醇脱盐和100%甲醇洗脱反相C8柱层析,获得硫苷脂纯品。最后,利用500YMC Diol液相色谱柱(3.0mm×250mm,5μm)分离,以正己烷/异丙醇(70∶30,v/v),异丙醇/水/甲酸/氨水(100∶13∶1∶0.14,v/v/v/v)为流动相梯度洗脱,采用负电喷雾电离(ESI)和母离子扫描模式,用液相色谱-串联质谱(LC-MS/MS)法分析了鳕鱼脑硫苷脂的纯度和分子种组成,并比较了其与哺乳动物脑中硫苷脂分子种组成的异同。结果表明,本实验制得的鳕鱼脑硫苷脂纯度为90.74%,与哺乳动物类似,鳕鱼脑硫苷脂的长链碱基以鞘氨醇为主,主要分子种为d18∶1-C24∶1,但脂肪酸的羟基化程度略低,且含少量独特的分子种,如d18∶1-22∶1和d18∶2-25∶2。  相似文献