共查询到20条相似文献，搜索用时 62 毫秒
1.
采用RT-PCR和RACE技术克隆鲫鱼slc7A8基因的全长cDNA序列,对基因序列用生物软件对基因进行生物信息分析,用Real-time PCR方法检测基因在组织中的mRNA表达丰度。结果表明,鲫鱼slc7A8cDNA全长为2709bp,包含195bp的5′UCR序列,921bp的3′UCR序列,1593bp开放阅读框,编码530个氨基酸序列;鲫鱼与斑马鱼基因同源性和编码氨基酸同源性分别为88.9%和95.5%,而与其他动物分别在70.1%-74.8%和80.6%-82.1%之间;预测编码蛋白的分子量为57719.84,等电点为4.8,对其结构预测显示该蛋白具有与哺乳动物十分相似的12个螺旋跨膜结构,跨膜区氨基酸高度保守,不同动物间的同源性都在92.2%以上;Real-time PCR检测鲫鱼组织中的mRNA表达丰度高低顺序为前肠、中肠、脑、后肠、肌肉、肾、鳃、心、肝脏。 相似文献
2.
利用RT-PCR和RACE方法克隆得到鳜鱼肝脏中控制高不饱和脂肪酸合成的脂肪酸去饱和酶(fatty acid desaturase,FAD)和脂肪酸延长酶(fatty acid elongase,ELO)基因的全长cDNA序列。FAD基因的全长cDNA序列1882bp,编码445个氨基酸,该蛋白序列含有FAD全部的特征结构区,包括3个组氨酸簇、2个跨膜区和1个细胞色素b5结构域,与其它鱼类FAD氨基酸序列的同源性为66.5%-90.1%,系统树分析显示其与欧洲鲈鱼和金头鲷等海水鱼类的亲缘关系最近。获得的ELO基因全长cDNA序列1401bp,编码294个氨基酸,该蛋白序列含有单一的氧化还原中心组氨酸簇、内质网停留信号和多个跨膜区等ELO特征结构,与其它几种鱼类ELO氨基酸序列的同源性为71.8%-86.7%,系统进化分析显示其与南方蓝鳍金枪鱼和金头鲷的亲缘关系较近。鳜鱼FAD和ELO基因全长cDNA序列的获得可为进一步研究其HUFA合成能力及调控机理奠定基础。 相似文献
3.
取正常泥蚶肌肉组织获得总RNA，纯化mRNA后，利用含SfiⅠ酶切位点的CDSⅢ引物逆转录合成cDNA第一链，通过LD—PCR合成cDNA双链，经胶回收除去短片段，与pDNR—LIB载体进行连接，电转化到大肠杆菌DH10B中，构建形成原始文库，进行滴度测试和重组率鉴定。结果表明原始文库的滴度为3．17×10^5cfu／ml，重组率为86．3％，插入片段多在500--2500bp间。随机挑选458个克隆测序，经分析获得94种已知基因及87种未知基因。其中包括铁结合蛋白（Ferritin）的全长cDNA序列。该基因序列全长895bp，5’-端非编码区为163bp，3’-非编码区为213bp，开放阅读框为519bp，编码172个氨基酸。推测其蛋白分子量和等电点分别为20kDa和4．89。氨基酸序列与皱纹盘鲍、太平洋牡蛎、血红扇头蜱、文昌鱼、中华蟾蜍、非洲爪蟾、小家鼠以及人等的同源性有62％-82％。比对结果表明，铁结合蛋白基因在动物进化中高度保守。 相似文献
5.
6.
大型多管水母的绿色荧光蛋白 总被引：3，自引：0，他引：3
采用PCR扩增和southern杂交筛选相结合的方法,从厦门水域的大型多管水母中分离到了新的绿色荧光蛋白基因gfpxm,并在大肠杆菌中进行了表达.gfpxmDNA序列编码区长为1042bp,包含3个外显子和两个内含子,cDNA编码区全长为717bp.gfpxmcDNA与已知野生型Aeqgfp 10cDNA长度一致,核苷酸同源性为81.9%,推导的氨基酸序列全长为238个氨基酸,与AeqGFP10的氨基酸同源性为83.6%.将gfpxm克隆至pTO-T7表达载体,GFPxm在大肠杆菌BL21中的表达量达菌体总蛋白的50%左右.荧光性质和强度分析结果表明,GFPxm蛋白的激发峰为476nm,发射峰为496nm,荧光量子产率为1. GFPxm蛋白的荧光很稳定,对热、碱性、变性剂和盐等有较强抗性. 相似文献
7.
利用RT-PCR和快速扩增cDNA末端(rapid amplification of cDNA ends,RACE)技术首次克隆了宽体沙鳅(Botia reevesae)-肌动蛋白基因的cDNA全序列,该序列全长为1795bp,由长100bp的5非翻译区(untranslated region,UTR)、570bp的3非翻译区和1125bp的开放阅读框(open reading frame,ORF)组成,编码375个氨基酸。宽体沙鳅-actin氨基酸序列包含1个糖基化位点、9个N-豆寇酰化位点、3个actin信号位点等主要结构区域。PSI-BLAST比对表明,宽体沙鳅-actin氨基酸与真鲷、罗非鱼、虹鳟等鱼类同源性达99%。NJ法系统进化分析显示宽体沙鳅-actin首先与鲢聚在一起,然后与、尖头等鱼类聚在一起。荧光定量PCR检测-actin基因在宽体沙鳅脑、鳃、心脏、肝、胃等12个组织的表达无显著差异(P<0.05),具有良好的稳定性。 相似文献
8.
海湾扇贝(Argopecten irradians)金属硫蛋白基因的克隆与分析 总被引：1，自引：1，他引：1
金属硫蛋白是一种普遍存在于生物体内的低分子量、半胱氨酸含量丰富、易于被外界刺激诱导的金属结合蛋白。采用表达序列标签法,结合cDNA末端快速扩增技术,首次获得了海湾扇贝金属硫蛋白(AiMT)的全长cDNA序列。该序列全长787bp,5′UTR(UntranslatedRegion)为79bp,3′UTR为270bp,开放阅读框(OpenReadingFrame,ORF)长度为438bp,可编码145个氨基酸。在其编码的氨基酸序列中半胱氨酸含量丰富,甘氨酸含量也较高,芳香族氨基酸含量低,不含组氨酸,存在有无脊椎动物和软体动物金属硫蛋白的特征序列CKCXXX-CXCX,C-末端的氨基酸序列也符合软体动物金属硫蛋白标签序列C-x-C-x(3)-C-T-G-x(3)-C-x-C-x(3)-C-x-C-K。序列特征分析表明,该序列具备金属硫蛋白的典型特征,是金属硫蛋白家族的成员。 相似文献
9.
半滑舌鳎的一种细胞凋亡抑制因子的克隆、鉴定与分析 总被引：1，自引：0，他引：1
克隆半滑舌鳎(Cynoglossus semilaevis)的1种细胞凋亡抑制因子survivin基因,并对其序列及表达模式进行分析.结果表明,该基因cDNA 序列全长704 bp(GenBank登录号为EU499314),包括86 bp5'非翻译区,444 bp开放阅读框以及含 poly(A)信号的174 bp的3'非翻译区,编码147个氨基酸.氨基酸序列同源性分析表明,半滑舌鳎survivin氨基酸序列与斑马鱼、非洲爪蟾、鸡、小鼠和人survivin氨基酸序列的同源性分别达到了63.3%,49.0%,43.7%,43.6%和41.5%.RT-PCR分析表明,survivin基因在半滑舌鳎成体雌鱼卵巢中大量表达,在雌鱼的肾脏中微量表达,在其它雌鱼的组织中无表达,在雄性的任何组织中均无表达.该基因在受精卵、多细胞和囊胚3个胚胎发育的早期阶段,大量表达,而随着胚胎发育的进程,survivin基因的表达逐渐减少,在孵化前尾芽期胚胎中只有微量表达,在孵化后5 d的仔鱼中,几乎检测不到survivin基因的表达.表明该基因可能参与卵子形成和胚胎的早期发育. 相似文献
10.
采用RT-PCR及RACE法从斜带石斑鱼Epinephelus coioides肝胰脏克隆得到胰蛋白酶原(trypsinogen,TRY)与淀粉酶(amylase,AMY)基因cDNA全序列.斜带石斑鱼肝胰脏TRY基因cDNA全长911 bp,其中5非翻译区(5'-UTR)为55bp,3'-UTR为127bp,开放阅读框(ORF)为729bp,编码242个氨基酸,包含所有丝氨酸蛋白酶中共有的高度保守的催化活性中心.序列一致性分析发现,斜带石斑鱼与牙鲆Paralichthys olivaceus、金头鲷Sparus aurata、鳎Solea senegalensis、石鲽Pleuronectes bicoloratus的TRY序列相似性高达86.8%-89.70%,与人Homo sapiens小鼠Mus musculus、斑马鱼Danio rerio的TRY相似性较低为59.9%-64.5%.斜带石斑鱼AMY基因cDNA全长1657bp,其中5'-UTR为41bp,3'-UTR为77 bp,ORF为1539bp,编码512个氨基酸,包含与哺乳动物α-AMY二级结构相似的8个α旋和8个β折叠.序列一致性分析发现,斜带石斑鱼与澳洲肺鱼Neoceratodus forsteri、美洲拟鲽Limanda americanus、大西洋鲑Salmo salar、斑马鱼AMY基因序列相似性高达82.4%-91.8%,与人、小鼠、鸡G.Gallus的AMY基因相似性较低为70.1%-72.3%.斜带石斑鱼TRY和AMY基因cDNA全序列的成功克隆为进一步研究其表达调控机理及研发有效提高其表达水平的饲料添加剂奠定基础. 相似文献
11.
Arne Schnack-Friedrichsen Angela M. Davis Jim D. Bennell Dei G. Huws 《Marine Georesources & Geotechnology》2001,19(4):221-243
The interdependence between the seismo-acoustic properties of a marine sediment and its geotechnical/physical parameters has been known for many years, and it has been postulated that this should allow the extraction of geotechnical information from seismic data. Though in the literature many correlations have been published for the surficial layer, there is a lack of information for greater sediment depths. In this article, a desktop study on a synthetic seafloor model illustrates how the application of published near-surface prediction equations to subsurface sediments (up to several tens of meters burial depth) can lead to spurious predictions. To test this further, acoustic and geotechnical properties were measured on a number of sediment core samples, some of which were subjected to loading in acoustically-equipped consolidation cells (oedometers) to simulate greater burial depth conditions. For low effective pressures (representing small burial depths extending to around 10 meters subsurface), the general applicability of established relationships was confirmed: the prediction of porosity, bulk density, and mean grain size from acoustic velocity and impedance appears generally possible for the investigated sedimentary environments. As effective pressure increases through, the observed relationships deviate more and more from the established ones for the near-surface area. For the samples tested in this study, in some instances increasing pressure even resulted in decreasing velocities. There are several possible explanations for this abnormal behavior, including the presence of gas, overconsolidation, or bimodal grain size distribution. The results indicate that an appropriate depth correction must be introduced into the published prediction equations in order to obtain reliable estimates of physical sediment properties for greater subsurface depths. 相似文献
12.
Shigehisa Nakamura 《Marine Geodesy》2000,23(1):55-61
Specific properties of the interannual sea level variations and annual tides in the Northwestern Pacific were studied. Several tide stations were monitored. The monthly mean sea level for the year of 1995 was analyzed at each tide station. A seismic event in 1995, some tectonic activity around the subject area, and the Kuroshio (the oceanic western boundary current) may possibly contaminate results which would have occurred from the astronomical annual tide alone. 相似文献
13.
The effect of different fixation and storage protocols on the flow cytometric (FCM) simultaneous analysis of bacterioplankton and phytoplankton in coastal seawater samples (Mediterranean coastal lagoons) was investigated. FCM measurements (cell number, fluorescence and scatter characteristics) were obtained through DAPI staining. Three fixatives [glutaraldehyde (GA), formaldehyde (FA) and paraformaldehyde (PFA)] and two storage (3 months duration) methods (5 °C and −196 °C) were tested. Two dominant populations were detected in studied samples: bacteria and eukaryotic picophytoplankton. Adding fixatives (2% final concentration) appears necessary to obtain FCM exhaustive counts of all the bacteria and phytoplanktonic cells. This was related to the permeation effect of fixatives which allowed a better DAPI staining of the cells. Maximum fluorescence, i.e. optimal staining of the cells was obtained with FA or PFA, and significant lower fluorescences with GA. Fixed samples stored at 5 °C induced rapid cell loss. Only storage in liquid nitrogen of samples fixed with FA or PFA, allows mid-term (≥4 months) preservation of bacteria or picophytoplankton cell numbers, and limited evolution of DAPI-induced fluorescence and scatter characteristics. 相似文献
14.
15.
《海洋学报(英文版)》2014,(9):F0003-F0003
<正>Acta Oceanologica Sinica(AOS)is a comprehensive academic journal edited by the Editorial Committee of Acta Oceanologica Sinica and is designed to provide a forum for important research papers of the marine scientific community which reflect the information on a worldwide basis.The journal publishes scholarly papers on marine science and technology,including physics,chemistry,biology, 相似文献
16.
The relationship between the flux of exotic benthic foraminiferal tests (i.e. tests which are supplied from open-sea sources alone) in a tidal inlet and that of bulk sediment was analysed, which can be expressed as two first-order linear equations. According to this relationship, in order to determine net sediment transport directions in the entrance, the test concentration in surficial sediments of the tidal basin can be compared against a ‘ critical level ’. The critical level is determined for the conditions that no net transport of bulk sediment is present within the entrance. If the observed concentration (averaged over the tidal basin) is higher than the simulated critical level, then the net sediment transport is directed to landward. This method is applied to the analysis of net sand transport at Christchurch Harbour, a tidal inlet system located in southern England. In this investigation, concentrations of exotic foraminiferal tests in the surficial sediments of the tidal basin and ebb tidal delta area were obtained from the analysis of sea-bed sediment samples. A series of probable critical levels were calculated based upon the data sets with regard to: (1) sediment discharge from the rivers; (2) magnitude of sediment discharge within the entrance during the ebb; (3) the test concentration outside the harbour; (4) the thickness of the moving layer; and (5) two parameters associated with dispersive processes. The results show that the concentration in the tidal basin sediment is higher than a number of simulated critical concentrations for representative cases. Consequently, the high level of the concentration of exotic benthic foraminiferal tests within the harbour should be explained as a result of landward net transport of sands within the entrance. 相似文献
17.
18.
The behaviour of an axially pre-loaded cylindrical member of an offshore structure, hit by a supply ship, has been investigated. The effects of axial pre-loading on the dynamic properties of members, the extent of damage and propagation of dynamic instability in the tubular members have also been investigated. Numerical models have been validated using available experimental data from the literature although these are mostly static with no pre-loading. Axial pre-loading was found to change the dynamic characteristics of some cylindrical members quite dramatically while it had no significant effect on others. The study examined whether there was any change in the behaviour between local and global modes when the loading was dynamic rather than static. The effect of damping on the dynamic instability of axially pre-loaded tubes under lateral dynamic loads was also studied. It was observed that for some geometries the quasi-static response can be used to define the boundary between bounded and unbounded dynamic responses. The main contribution of this study, compared with previous investigations reported in the literature, is that both pre-loading and dynamic effects have been included. 相似文献
19.
Rahul Sharma 《Marine Georesources & Geotechnology》2000,18(3):237-250
Observations on seafloor features from subbottom profiling and seabed photography data show that the undisturbed and natural conditions before the benthic disturbance undergo a substantial change after operation of the disturber. The effects can be seen in the form of deep trenches and grooves, formed by the action of the sleds and the pumping of the sediment along its path, as well as sediment piles on either side of the track. Biological traces such as fecal coils, casts, tubes, burrows, and trails get obliterated in the areas of disturbance and resedimentation. The smooth, uniform nature of the seafloor observed in predisturbance phase shows microtopographic changes in and around the disturbed area as a result of sediment excavation and resettlement. Vertical mixing as well as lateral transport of sediment alter the geological, biological, physical, and chemical conditions on the seafloor, which need to be monitored over time to assess the process and the time taken for restoration of environmental conditions. 相似文献
20.
Vance P. Vicente 《Marine Ecology》1989,10(2):179-191
Abstract. Long-term changes in the distributional patterns of commercial sponges ( Spongia spp. and Hippospongia spp.) within the West Indian Region indicates that: 1) commercial sponges had a widespread distribution throughout the whole West Indian Region and were ubiquitous in very shallow water until about the first half of the present century; 2) they were fished commercially not only in the traditional northern Caribbean sites (Florida, Gulf of Mexico, Bahamas) but also in the Greater ( e. g. , Hispaniola, Jamaica) and Lesser Antilles; and 3) they became extinct throughout most of the Lesser Antilles ( e. g. , Puerto Rico, Vieques, St. Thomas) sometime during the first half of this century. Mortalities of spongiids within the Antilles were found to differ from other marine mortalities in that: 1) species disappeared from a large region; 2) species vanished from different habitats and depths; and 3) natural populations never recovered. Species richness patterns suggest that commercial sponge genera ( Spongia and Hippospongia ) evolved under slightly cooler elimatic conditions than those found at present, and that these extinctions occurred as a direct or indirect effect of positive thermal anomalies in sea surface and atmospheric temperatures between 1900–1950. The concept that species diversity is stable on a regional scale is questioned. 相似文献