DNA identification of Pacific bluefin tuna (Thunnus orientalis) in the New Zealand fishery     

P. J. Smith  L. Griggs  S. Chow 《新西兰海洋与淡水研究杂志》2013,47(4):843-850

Muscle samples were collected from 69 specimens identified as Pacific bluefin tuna (Thunnus orientalis) (Temminck and Schlegel, 1844) in the New Zealand Exclusive Economic Zone (EEZ) between 1990 and 2000. Identifications before 1996 were based on body size and colour of the caudal keel; later identifications were mostly based on the shape of abdominal cavity. The tissue samples were tested with a diagnostic mitochondrial DNA marker that distinguishes southern bluefin Thunnus maccoyii (Castelnau, 1872) and Pacific bluefin tuna T. orientalis; 59 specimens were confirmed as T. orientalis and 10 as T. maccoyii. Specimens recorded as Pacific bluefin tuna by the shape of the abdominal cavity were correctly identified as T. orientalis, and this character can be used to identify large specimens landed on tuna vessels. Some specimens recorded as Pacific bluefin tuna on the basis of colour and size were T. maccoyii; and early records of T. orientalis in New Zealand waters, based on these characters, are unreliable. Unusual colour patterns were reported in some specimens of T. orientalis but not T. maccoyii. The Pacific bluefin tuna T. orientalis accounted for less than 0.3% of the bluefin tuna catch in the New Zealand EEZ during the 1990s.  相似文献   

草鱼出血病病毒VP7蛋白基因的人工合成与原核表达   总被引：2，自引：0，他引：2  

韦先超  杨泽晓  王印  裴腊梅  姚学萍  汪开毓  杨水仙 《海洋与湖沼》2013,44(3):645-650

根据GenBank已发表的GCRVVP7蛋白基因序列,设计合成2条特异性引物和19条搭桥引物,通过搭桥PCR人工合成带有3处变异碱基的GCRVVP7蛋白基因整个编码框,经过酶切、连接、PCR鉴定和测序鉴定克隆到原核表达载体pColdTF中,并转化大肠杆菌BL21(DE3),进行IPTG诱导表达和表达产物的SDS-PAGE、Western-blotting分析、纯化以及反应原性的iELISA检测。结果表明,成功人工合成了长831bp的GCRV VP7蛋白基因编码框和构建了VP7蛋白基因的重组质粒pCold TF-VP7,pColdTF-VP7转化菌经IPTG(1mmol/L)诱导3—4h即可获得特异性蛋白VP7重组蛋白的高效表达,VP7重组蛋白相对分子量大小约为73.9ku,与预期大小相符,且与鼠抗GCRV血清有良好反应原性。  相似文献   

泥蚶(Tegillarca granosa)生长因子受体结合蛋白2(GRB2)基因的克隆与表达分析   总被引：3，自引：0，他引：3  

董迎辉  项翔  姚韩韩  包永波  孙长森  林志华 《海洋与湖沼》2013,44(4):937-943

通过已构建的泥蚶(Tegillarca granosa)转录组文库, 利用SMART RACE技术扩增得到泥蚶Tg-GRB2基因的全长cDNA序列, 并对其生物信息学、组织表达及发育阶段表达特征进行了分析。结果表明, 泥蚶Tg-GRB2 cDNA全长1283bp, 开放阅读框为708bp, 编码236个氨基酸; 蛋白结构预测显示, Tg-GRB2蛋白包含SH3-SH2-SH3三个功能域, SH2结构域由两端的α-螺旋和中间反向平行的β-折叠片构成, SH3结构域主要由β-折叠片组成, 具备典型的结构特征; 氨基酸序列比对发现, Tg-GRB2与脊椎动物的同源性为62.1%—63.8%, 而与玻璃海鞘和日本血吸虫同源性较低。qRT-PCR检测结果显示: Tg-GRB2 mRNA在泥蚶血液、斧足、鳃、外套膜、闭壳肌、内脏团6个组织中都有表达, 而在血液中的表达量极显著地高于其它组织; 在泥蚶的不同发育阶段中, Tg-GRB2 mRNA在 2—4细胞胚胎、原肠胚、担轮幼虫、D形幼虫中均有较高的表达量, 而在D形幼虫中表达量最高, 表明Tg-GRB2基因在泥蚶早期发育中发挥重要调节作用。  相似文献   

基于马氏珠母贝(Pinctada fucata)血细胞全长转录组数据补体样组分的鉴定与分析     

王菁  李桂英  林楷琪  欧静  张淑瓶  王忠良 《海洋与湖沼》2022,53(1):176-186

补体系统作为先天免疫的重要组成部分,是一种复杂的限制性蛋白水解系统,其在免疫系统中发挥着重要的防御作用。为分析马氏珠母贝补体系统的组成及作用机制,使用血细胞样品进行了全长转录组测序建库、基因比对、功能注释,共挖掘到212个潜在补体样组分相关基因。补体样组分基因经同源性比对和结构域检测分析表明,检索到的基因分别编码89个含C1q结构域蛋白、57个C型凝集素蛋白、33个纤维胶凝蛋白、11个纤维蛋白原相关蛋白、8个甘露糖结合型凝集素关联丝氨酸蛋白酶、2个含硫酯蛋白(1个C3分子,1个TEP分子)、1个补体受体、2个补体因子、9个丝氨酸蛋白酶。随机选择12个补体相关基因,使用溶藻弧菌刺激前后的血细胞样品进行实时定量PCR检测其表达水平,结果显示C1q(C1q domain containing protein)、C-lectin、MBL(mannose-binding lectin)、ficolin、MASP(mannan-binding lectin serine protease)等基因均呈现出显著差异表达,表明马氏珠母贝补体系统是一个复杂的多组分效应系统,且可能通过凝集素途径或类似于凝集素途径激活补体系统的免疫作用。研究结果为进一步验证马氏珠母贝中存在的原始补体系统提供了分子生物学证据,同时对深入了解马氏珠母贝免疫防御机制,丰富和发展海洋无脊椎动物免疫学内容也具有重要理论意义。  相似文献   

青蛤(Cyclina sinensis)TLR2基因的克隆与表达分析   总被引：2，自引：2，他引：0  

任毅鹏  高晶  潘宝平  高虹  闫春财 《海洋与湖沼》2014,45(5):1037-1043

利用转录组文库分析得到青蛤(Cyclina sinensis)的Toll样受体2(Toll-like receptor 2,TLR2)基因序列,采用荧光定量PCR法分析该基因的表达过程。结果显示,青蛤TLR2基因的cDNA开放阅读框为2082bp,编码693个氨基酸,具典型的LRRs、单次跨膜区和TIR结构域。该基因在血淋巴中的表达量最高,与肝脏、鳃、外套膜、闭壳肌和性腺有显著性差异(P0.05)。在鳗弧菌和Poly I:C刺激下,青蛤血淋巴中TLR2基因3h开始升高,48h达到最大值,与对照组和空白组有极显著性差异(P0.01);藤黄微球菌刺激下,血淋巴中CsTLR2基因3h开始升高,48h亦达到最大值,实验组与对照组有显著性差异(P0.05)。  相似文献   

凡纳滨对虾(Litopenaeus vannamei)Runt转录因子的基因克隆及其功能分析     

王晓雯  梁艳  邢婧  刘旭雅  黄倢 《海洋与湖沼》2014,45(6):1338-1345

Runt结构域转录因子家族(Runx)在哺乳动物体内存在三种基因型,分别起着造血,骨生成和控制上皮细胞增殖的作用。本文应用反转录和c DNA末端快速扩增(RACE)技术,首次在凡纳滨对虾(Litopenaeus vannamei)中克隆并测序了一种Runt(lvrunt)基因全长。lvrunt c DNA全长1754bp,含有1个663bp长的开放阅读框,编码221个氨基酸,理论分子量为23.6k Da,具有Runt-family结构域。对凡纳滨对虾鳃、肠、肝胰腺、类淋巴、心脏、肌肉组织和血淋巴lvrunt的转录特征进行分析,结果显示该基因几乎特异性地在血淋巴细胞中表达,而在其他组织中表达较低。肌肉注射重组造血激素蛋白(r AST)或白斑综合征病毒(WSSV)粗提液均可显著提高lvrunt在凡纳滨对虾体内的转录表达。上述结果提示,lvrunt主要在血细胞发挥作用,可能作为造血激素的下游基因起到调节血细胞增殖和分化的作用,并在受到病毒感染后,参与提升血细胞数量或者促进血细胞分化的过程。  相似文献   

自动气象站数据采集系统的传感器接口     

陶伟  姚振东 《成都信息工程学院学报》2006,21(Z1):53-58

自动气象站观测系统在气象领域中应用广泛,数据采集器是系统中的关键部件,改进了低功耗、多功能、多通道、智能化控制等性能.传感器接口包括DAC实现的可调恒压源和可调恒流源、稳压二极管实现的前端保护、多路模拟开关实现的通道控制和测量类型控制、精密线性光耦合器实现的隔离电路、DAC和仪表放大器实现的增益控制等硬件部件.设计单片机程序实现电路的各种控制及要求功能.经制板、调试,该数据采集器能正常运行.  相似文献   

Fast tidal cycling and the origin of life     

Richard Lathe 《Icarus》2004,168(1):18-22

Replicating prebiotic polymers are thought to predate the emergence of true life-forms. The initial mode of replication, a prerequisite for Darwinian selection, is unknown, but demands an explanation based on local physicochemistry. Dual consideration of the conditions of the early terrestrial surface, with the unusual physicochemical properties of nucleic acids like DNA, could explain the emergence of nucleic acids as key biomolecules. The early impact that produced the Moon, and fast terrestrial rotation, subjected coastal areas 3.9 Ga ago to rapid tidal flooding (dilution) and drying (concentration), with a likely periodicity in the range of 2-6 h, and could have provided a driving force for cyclic replication of early biomolecules. Such a mechanism applies only to molecules capable of association/polymerization at high salt concentration, and of dissociation at low salinity. Nucleic acids meet these criteria. It is suggested that tidal cycling, resembling the polymerase chain reaction (PCR) mechanism, could only replicate and amplify DNA-like polymers. This mechanism suggests constraints on the evolution of extra-terrestrial life.  相似文献   

Transformation of ammonium nitrogen and response characteristics of nitrifying functional genes in tannery sludge contaminated soil          下载免费PDF全文

Kong Xiang-ke  Zhang Zi-xuan  Wang Ping  Wang Yan-yan  Zhang Zhao-ji  Han Zhan-tao  Ma Li-sha 《地下水科学与工程》2022,10(3):223-232

High concentrations of ammonium nitrogen released from tannery sludge during storage in open air may cause nitrogen pollution to soil and groundwater. To study the transformation mechanism of NH₄⁺-N by nitrifying functional bacteria in tannery sludge contaminated soils, a series of contaminated soil culture experiments were conducted in this study. The contents of ammonium nitrogen (as NH₄⁺-N), nitrite nitrogen (as NO₂^?-N) and nitrate nitrogen (as NO₃^?-N) were analyzed during the culture period under different conditions of pollution load, soil particle and redox environment. Sigmodial equation was used to interpret the change of NO₃^?-N with time in contaminated soils. The abundance variations of nitrifying functional genes (amoA and nxrA) were also detected using the real-time quantitative fluorescence PCR method. The results show that the nitrification of NH₄⁺-N was aggravated in the contaminated silt soil and fine sand under the condition of lower pollution load, finer particle size and more oxidizing environment. The sigmodial equation well fitted the dynamic accumulation curve of the NO₃^?-N content in the tannery sludge contaminated soils. The Cr(III) content increased with increasing pollution load, which inhibited the reproduction and activity of nitrifying bacteria in the soils, especially in coarse-grained soil. The accumulation of NO₂^?-N contents became more obvious with the increase of pollution load in the fine sand, and only 41.5% of the NH₄⁺-N was transformed to NO₃^?-N. The redox environment was the main factor affecting nitrification process in the soil. Compared to the aerobic soil environment, the transformation of NH₄⁺-N was significantly inhibited under anaerobic incubation condition, and the NO₃^?-N contents decreased by 37.2%, 61.9% and 91.9% under low, medium and high pollution loads, respectively. Nitrification was stronger in the silt soil since its copy number of amoA and nxrA genes was two times larger than that of fine sand. Moreover, the copy numbers of amoA and nxrA genes in the silt soil under the aerobic environment were 2.7 times and 2.2 times larger than those in the anaerobic environment. The abundance changes of the amoA and nxrA functional genes have a positive correlation with the nitrification intensity in the tannery sludge-contaminated soil.  相似文献   

A Sensitive and Versatile Multiplex PCR System for the Rapid Detection of Enterotoxigenic (ETEC), Enterohaemorrhagic (EHEC) and Enteropathogenic (EPEC) Strains of Escherichia coli     

R.Y.C Kong  C.L So  W.F Law  R.S.S Wu 《Marine pollution bulletin》1999,38(12):21-1215

Although Escherichia coli is widely distributed in the marine environment, only a small percentage are pathogenic to humans. Nonetheless, the widespread occurrence of waterborne infections of E. coli origin in humans has become one of the major health problems worldwide. To date, several types of enterovirulent E. coli have been recognized as the aetiologic agents of various gastrointestinal infections in humans. The most commonly encountered are those belonging to the enterotoxigenic (ETEC), enteroinvasive (EIEC), enterohaemorrhagic (EHEC) and enteropathogenic (EPEC) subtypes. In order to better determine the health risks that are associated with exposure to some of these specific subtypes, we have developed a very sensitive multiplex PCR system for the rapid detection and typing of ETEC, EHEC and possibly EPEC strains of E. coli in the aquatic environment. The target genes chosen for this investigation included: the PHO-A housekeeping gene (present in all E. coli); the LT1, LT2 and ST1 genes of ETEC; the VT1 and VT2 verotoxin, and EAE virulence genes of EHEC and EPEC, respectively. Six pairs of oligonucleotide primers were designed to simultaneously amplify internal fragments of these genes by multiplex PCR to generate PCR products that could be analysed and confirmed with relative ease by gel electrophoresis and HincII enzyme digestion. The results showed that the six sets of PCR primers were highly specific for their target genes and produced specific amplimers of the expected size from several control strains of E. coli – ATCC 35401 (LT1+/ST1+); SA53 (LT2+/VT2+); and O157 (VT1+/VT2+/EAE+). The detection sensitivity of the multiplex PCR system for the six target genes in an E. coli cell mixture was optimized and enhanced by preincubating serially diluted cells in Luria-Bertani broth for 6 h prior to PCR analysis. The results obtained indicated a detection sensitivity of 10° CFU (of each strain) per 100 μl reaction. Multiplex PCR analysis of seawater samples collected from four sewage-polluted sites in Hong Kong indicated the presence in all four samples of E. coli bacteria that were positive for LT1, ST1, VT1 and EAE virulence genes. Overall, the data indicated that the multiplex PCR system described in this study is a potentially very useful and powerful method for routine monitoring and risk assessment of water quality.  相似文献