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1.
《渔业科学进展》2016,(6)
本研究利用RACE和RT-PCR技术克隆了海蜇(Rhopilema esculentum)磷脂酶A_2基因(Re-PLA_2-1)的cDNA及基因组序列,并分析了其m RNA在海蜇不同发育阶段的表达。Re-PLA_2-1基因的cDNA全长为824 bp,包括了48 bp的5?非编码区、504 bp的开放阅读框及272 bp的3'非翻译区。SMART分析显示,Re-PLA_2-1为分泌蛋白,包括了一个由19个氨基酸组成的信号肽和一个由118个氨基酸组成的磷脂酶A_2结构域。多序列比对和系统进化分析显示,Re-PLA_2-1基因与来自星状海葵(Nematostella vectensis)、僧袍芋螺(Conus magus)、长牡蛎(Crassostrea gigas)等磷脂酶A_2的相似性较高,共同聚类为pfam09056 GIX PLA_2分支,均包含pfam09056家族成员酶活性所必需的Ca~(2+)结合位点、活性催化位点和PLA_2结构域所必需形成二硫键的半胱氨酸。Re-PLA_2-1基因组全长为2671 bp,由4个外显子和3个内含子组成。RT-PCR结果显示,Re-PLA_2-1基因在海蜇4个发育阶段均有表达,其中,横裂体阶段的表达量最高,碟状体阶段最低。这些研究结果为进一步了解海蜇磷脂酶A_2毒素的生物功能奠定了基础。 相似文献
2.
基于转录组454 GS FLX测序结果,利用RACE和RT-PCR技术克隆了海蜇(Rhopilema esculentum) Notch基因的cDNA全长,并分析了其mRNA在海蜇不同发育阶段的表达差异,以探讨Notch基因对海蜇无性生殖的影响.结果显示,海蜇Notch基因的cDNA全长为6768 bp,包括90 bp的5'非编码区、6066 bp的开放阅读框及612 bp包含AATAAA加尾信号的3'非翻译区.SMART分析表明,海蜇Notch为分泌蛋白,其信号肽由21个氨基酸组成.成熟肽由2000个氨基酸组成,包括37个结构域、26个表皮生长因子样结构域、3个富含半胱氨酸的Notch/Lin-12(NL)结构域、1个NOD结构域、1个NODP结构域、1个跨膜结构域和6个锚蛋白结构域ANK,并含有25个N-糖基化位点.同源分析表明,海蜇Notch与来自刺胞动物门的海葵(Nematostella vectensis)的Notch相似性为39％,与无脊椎动物和脊椎动物的氨基酸相似度分别为35％-37％和34％-38％.RT-PCR分析表明,Notch基因在海蜇无性繁殖的4个发育时期均有表达,螅状体阶段的表达量最高,横裂体阶段表达量最低,螅状体的表达量是横裂体的1.85倍.这些研究结果为进一步研究Notch信号通路在海蜇无性繁殖中的调控作用奠定了基础. 相似文献
3.
黄鳝PLA2基因的克隆及表达分析 总被引：1，自引：0，他引：1
利用同源克隆技术和RACE技术克隆黄鳝(Monopterus albus)磷脂酶A2(PLA2)基因的cDNA全长;并采用荧光定量RT-PCR法检测PLA2基因在黄鳝各组织中的表达量情况。结果显示:黄鳝PLA2基因全长cDNA大小为2 606 bp(Gen Bank登录号:KX852397),其中开放阅读框2 205 bp,编码736个氨基酸,预测分子大小为83.623 kD,理论等电点5.84;5'和3'非编码区长度分别为208 bp和193 bp。该蛋白序列没有信号肽和跨膜结构。氨基酸序列比对和系统树分析显示,黄鳝PLA2基因的结构十分保守,黄鳝与大黄鱼和金头鲷的PLA2基因亲缘关系最近,与鼠类和蟾蜍的亲缘关系较远。RT-PCR分析表明,PLA2基因在组织中的表达具有组织特异性,在消化器官前肠组织中的表达最高,在性腺及肌肉组织中表达很少。 相似文献
4.
《渔业科学进展》2017,(4)
基于转录组454 GS FLX测序结果,利用RACE和RT-PCR技术克隆了海蜇(Rhopilema esculentum)Notch基因的c DNA全长,并分析了其m RNA在海蜇不同发育阶段的表达差异,以探讨Notch基因对海蜇无性生殖的影响。结果显示,海蜇Notch基因的c DNA全长为6768 bp,包括90 bp的5?非编码区、6066 bp的开放阅读框及612 bp包含AATAAA加尾信号的3?非翻译区。SMART分析表明,海蜇Notch为分泌蛋白,其信号肽由21个氨基酸组成。成熟肽由2000个氨基酸组成,包括37个结构域、26个表皮生长因子样结构域、3个富含半胱氨酸的Notch/Lin-12(NL)结构域、1个NOD结构域、1个NODP结构域、1个跨膜结构域和6个锚蛋白结构域ANK,并含有25个N-糖基化位点。同源分析表明,海蜇Notch与来自刺胞动物门的海葵(Nematostella vectensis)的Notch相似性为39%,与无脊椎动物和脊椎动物的氨基酸相似度分别为35%–37%和34%–38%。RT-PCR分析表明,Notch基因在海蜇无性繁殖的4个发育时期均有表达,螅状体阶段的表达量最高,横裂体阶段表达量最低,螅状体的表达量是横裂体的1.85倍。这些研究结果为进一步研究Notch信号通路在海蜇无性繁殖中的调控作用奠定了基础。 相似文献
5.
通过同源克隆和RACE技术获得斑节对虾(Penaeus monodon)亲环素A(cyclophilin A,CYPA)的cDNA序列.根据已克隆的cDNA序列设计引物,利用染色体步移技术(Cenomic DNA walking)从斑节对虾卵巢组织中克隆了CYPA基因的启动子和基因组DNA序列.测序结果表明,斑节对虾亲环素A(PmCYPA)cDNA全长834 bp,其中开放阅读框(open reading frame,ORF)为495 bp,可编码164个氨基酸;5'非编码区为31 bp,3'非编码区为308 bp.从斑节对虾基因组文库中扩增的CYPA基因组DNA全长3 181 bp,其中包括启动子区域1 173 bp,1个内含子426 bp,2个外显子序列:分别为101 bp和399bp.在5'UTR上游区域有明显的启动子序列,包含一个GC盒和2个CAAT盒,同时还包含AP1、CRE等调控元件,符合真核生物典型的启动子特征.分析基因的结构表明所克隆的PmCYPA符合PPlase家族成员特征,属于此基因家族成员. 相似文献
6.
采用RACE技术解析了海蜇(Rhopilema esculentum)Frizzled1基因的cDNA和基因组结构:Re-Fzd1基因的全长cDNA为2387 bp,其中编码区为1761bp,编码586个氨基酸的多肽。SMART分析表明,Re-Fzd1基因具备Fzd家族共同的结构特征,包括:一个由23个氨基酸组成的信号肽,一个位于N-末端富含10个保守半胱氨酸残基的半胱氨酸富集域(CRD),一个含有7个跨膜片段的跨膜结构域,以及一个含有5个重要的磷酸化位点的C端尾巴。多序列比对表明,Re-Fzd1基因与刺胞动物贝螅(Hydra echinata)、水螅(Hydra vulgaris)、半球美螅水母(Clytia hemisphaerica)和海葵(Nematostella vectensis)Fzd1具有高度相似性,与来自脊椎动物人(Homo sapiens)、鼠(Mus musculus)、爪蟾(Xenopus laevis)和斑马鱼(Danio rerio)的Fzd1、Fzd2和Fzd7家族基因也具有较高的同源性。基于N-J法,将人、鼠、爪蟾、斑马鱼和果蝇(Drosophila melanogaster)所有Fzd家族基因系统进化分析显示,除果蝇外,所有Fzd家族成员聚类成4个类群,11个亚家族,海蜇Re-Fzd1基因首先与刺胞动物门的Fzd1聚类在一起,然后与脊椎动物Fzd1、Fzd2和Fzd7三个家族聚成一个类群,表明脊椎动物Fzd1、Fzd2和Fzd7家族可能与刺胞动物门的Fzd1起源于同一个共同祖先。Re-Fzd1基因组序列中不含有内含子。实时荧光定量PCR结果显示,Re-Fzd1基因在海蜇无性繁殖的4个发育阶段均有表达,其中,表达量最高的横裂体阶段是表达量最低的稚水母阶段的3.67倍。整体原位杂交显示,在海蜇横裂体时期,Re-Fzd1原位表达在触手、基座及发生横裂的部位。这些结果都表明,Re-Fzd1不但参与了海蜇的早期发育过程,还调控了海蜇无性繁殖的发生。 相似文献
7.
鳜脂蛋白脂酶和肝脂酶基因结构与组织表达 总被引：3，自引：0，他引：3
为了研究鳜(Sinioerca chuatsi)脂蛋白脂酶(LPL)、肝脂酶(HL)基因结构与功能关系,首先采用RT-PCR及RACE法,克隆得到鳜肝脏LPL与HL基因cDNA全序列.鳜肝脏LPL基因cDNA全长为2089 bp,其中开放阅读框(ORF)为1 548 bp,编码515个氨基酸.鳜HL基因cDNA全长为1 964 bp,其中ORF为1 494 bp,编码497个氨基酸.进化树分析显示,鳜LPL与HL基因与其他脊椎动物的LPL、HL基因各自聚集成簇.对鳜基因组进行PCR获得鳜LPL与HL全基因组DNA序列,与其他脊椎动物基因结构相似,鳜LPL基因由10个外显子和9个内含子组成,全长7 392 bp;鳜HL基因南9个外显子和8个内含子组成,全长8 837 bp.应用Genome Walker方法在鳜克隆得到一段长为1 071 bp的LPL和一段长为2 173 bp的HL基因5'侧翼Ⅸ序列,并利用相关软件预测其中具有多个保守的顺式调控元件.组织表达研究显示与易患动脉粥样硬化的人类LPL不同,鳜LPL基因在所有被检测组织中均有表达:在脂肪组织中大量表达,其次是肝脏、脑、肠道、肌肉,在脾中表达量最低;而鳜HL基因的表达则与人类相似,只在肝脏中表达.本研究将为深入探讨机体脂质代谢调节机制以及LPL、HL在防治动脉粥样硬化中的作用奠定良好基础. 相似文献
8.
牙鲆dazl基因的克隆与表达分析 总被引：1，自引：0，他引：1
dazl(deleted in azoospermia-like)基因是多种物种精子发生的重要调控因子,为探讨该基因在牙鲆(Paralichthys olivaceus)性腺发育分化中的作用,本研究采用同源克隆和RACE技术克隆了牙鲆dazl基因的cDNA序列,利用生物信息学方法分析了其序列结构,并利用实时荧光定量PCR技术检测了该基因在牙鲆成体组织和早期发育阶段的表达情况。结果发现:牙鲆dazl cDNA序列全长2 031 bp,包括105 bp的5'端非翻译区,1 275 bp的3'端非翻译区和编码217个氨基酸残基的651 bp开放阅读框;氨基酸同源序列比对显示该序列存在一个高度保守的RRM(RNA recognition motif)结构域,且该结构域在鱼类中具有100%的序列一致性;荧光定量PCR结果显示dazl基因主要在牙鲆性腺中表达,且在精巢中的表达高于卵巢;在牙鲆早期发育阶段,dazl基因在未受精卵、受精卵及囊胚期均有较高的转录本,而在原肠胚期表达开始降低,至孵化后3 d一直保持较低的水平,这些丰富表达的母源性dazl mRNA可能参与了胚胎发育中原始生殖细胞的形成。本研究为进一步阐明dazl基因在牙鲆生殖发育中的功能奠定了基础。 相似文献
9.
根据本实验室前期获得的脊尾白虾(Exopalaemon carinicauda)α2-巨球蛋白基因EST序列,采用cDNA末端快速扩增(Rapid amplification of cDNA end, RACE)技术克隆获得脊尾白虾α2-巨球蛋白基因cDNA全长,命名为Ecα2M基因.该基因全长4823 bp,由4413 bp的开放阅读框、64 bp的5'端非编码区以及346 bp的3'端非编码区组成.开放阅读框编码1470个氨基酸,分子量为163.0 kDa,理论等电点为5.03.序列分析显示,Ecα2M序列N端含有23个氨基酸组成的信号肽.同源性分析显示,脊尾白虾Ecα2M氨基酸序列与罗氏沼虾(Macrobrachium rosenbergii)α2M的同源性最高,达到80%.荧光定量PCR分析结果显示,Ecα2M基因在血细胞、肝胰腺、肌肉、鳃、卵巢、眼柄、胃及肠中均有表达,其中在血细胞中的相对表达量最高.感染鳗弧菌和WSSV后,脊尾白虾血细胞中Ecα2M的相对表达量于6 h达到最大值且显著高于对照组(P＜0.05),肝胰腺中Ecα2M的相对表达量于3 h达到最大值且显著高于对照组(P＜0.05),相对表达量变化具有明显的时间差异性. 相似文献
10.
黄颡鱼LRH-1基因的分离和表达 总被引：1，自引：0，他引：1
肝受体类似物(Liver receptor homolog-1,LRH-1)是核受体家族Ftz-F1中一个亚家族成员,由于其在哺乳类卵巢中的高表达而被认为在卵巢类固醇合成的调控中起着一定的作用.本研究使用RT-PCR、RACE法分离到了黄颡鱼(Pelteobagrus fulvidraco)肝受体类似物基因(LRH-1)以及一个无活性的LRH-1' cDNA.LRH-1 cDNA全长1 945 bp,包括γ'非翻译区207bp,3'非翻译区238 bp,开放阅读框1 500 bp,编码500个氨基酸,推算的蛋白分子量为57.19 kD.LRH-1'全长1 663 bp,5'非翻译区283 bp,3'非翻译区330 bp,开放阅读框1 050 bp,编码350个氨基酸,推算的蛋白分子量为39.7 kD.与LRH-1相比,LRH-1'缺少存在于基因5'端的锌指结构和3'端起转录激活作用的AF-2基序.不同动物LRH-1氨基酸序列比对结果表明,黄颡鱼LRH-1和其他动物的LRH-1相似性为79%～85%,和其他动物的类固醇生成因子(SF-1)相似性为59%～66%.系统发育树显示,Ftz-F1分为两大支,一支包括鱼类SF-1;另一支又有2个分支,分别包含其他动物的SF-1和所有动物的LRH-1,在LRH-1分支中鱼类LRH-1单独为一小支.以β-actin为内标的荧光实时定量.RT-PCR结果显示,LRH-1在雌、雄黄颡鱼脑、肝、肾、性腺均有表达,以肝脏的表达量为最高,雌鱼各组织的表达量均比雄鱼高,其中肝脏和性腺差异明显(P＜0.05);在卵巢的不同发育阶段,该基因的表达量也不尽相同,Ⅲ期卵巢表达量最高,明显高于Ⅴ期(P＜0.05),Ⅴ期又明显高于Ⅱ、Ⅳ和Ⅵ期(P＜0.05);注射催产素促黄体释放激素类似物(LRH-A)12 h后,LRH-1在脑的表达量没明显变化,在肝脏和肾脏的表达量明显增加,而在卵巢的表达量明显下降. 相似文献
11.
《Journal Of Aquatic Food Product Technology》2013,22(4):61-70
No abstract available for this article. 相似文献
12.
Shun Watanabe Jun Aoyama Seishi Hagihara Bunpei Ai Rhodora V. Azanza Katsumi Tsukamoto 《Fisheries Science》2013,79(3):375-383
Anguilla luzonensis and A. huangi were each described in 2009 using eels obtained from northern Luzon Island. We examined the taxonomic status of these two groups of eels using morphological and molecular genetic characters. There were no significant differences in two vertebrae counts between eels of A. luzonensis and A. huangi. Mitochondrial 16S ribosomal RNA and cytochrome b genes sequences were obtained and compared among 28 specimens of A. luzonensis, the holotypes of A. luzonensis and A. huangi, and one specimen of the other 15 anguillid species. The specimens of A. luzonensis exhibited almost identical sequences, including the holotype, with only a few site differences, and the genetic difference between the holotypes of A. luzonensis and A. huangi was within the range of differences of specimens of A. luzonensis. The other anguillid species were genetically very different from A. luzonensis and A. huangi, although A. interioris is a closely related species. It is clear that A. luzonensis and A. huangi are the same species, and according to the principle of priority in zoological nomenclature, A. luzonensis Watanabe, Aoyama, and Tsukamoto, 2009 is the valid species name, and A. huangi Teng, Lin, and Tzeng, 2009 is a junior synonym of A. luzonensis. 相似文献
13.
The effects of exposing the eggs of Pacific threadfin and amberjack eggs (AEs) to different concentrations of hydrogen peroxide for 5 min on hatch rate and survival were assessed in a series of experiments using a petri dish model rearing system. Despite significant inter‐batch variation in hatch rate, it was shown that eggs of both species could be safely exposed to up to 11 340 mg L−1 H2O2 for 5 min. Exposure to 34 230 mg L−1 H2O2 for 5 min was shown to be lethal to AEs at a late stage of development. In two further experiments, it was demonstrated that Pacific threadfin eggs were resistant to all tested concentrations of a range of polyvinylpyrrolidone iodine (PVP‐I) concentrations and contact times (up to 1000 mg L−1 PVP‐I for 10 min). The level of bacteria adhering to the eggs of both species was highly variable. Where eggs were heavily colonized (>104 cfu egg−1), hydrogen peroxide concentrations of at least 11 340 mg L−1, or PVP‐I concentrations higher than 500 mg L−1 for 10 min, were required for effective sterilization. In less colonized batches, rinsing in sterile seawater or exposure to lower (550 mg L−1) concentrations of H2O2 was sufficient to result in high apparent levels of surface sterility (<1 cfu egg−1). 相似文献
14.
Mulloway (Argyrosomus japonicus) occur in estuarine and coastal waters surrounding Australia, Africa, India, Pakistan, China, Korea and Japan, where they are important in fisheries. This study identified that mulloway in south-eastern Australia had similar growth rates, but matured at smaller lengths and younger ages, to those in South Africa and Western Australia. Growth of both sexes was similar to about 5 years, after which females grew faster and attained a greater maximum length than males. Female mulloway matured at 4–5 years of age with a L50 of 68 cm, whereas males matured at 2–3 years of age with a L50 of 51 cm. The commercial fishery in New South Wales was characterised by declining catches and a reduction in the proportion of mulloway of mature lengths in landings. During 2002–2005 commercial landings were dominated (83%) by fish within 15 cm of the current minimum legal total length of 45 cm and aged 2 and 3 years (>80%), even though mulloway can attain lengths of 200 cm and live >30 years. Estimates of the rates of instantaneous total mortality ranged between 0.34 and 0.45, whilst the rate of instantaneous natural mortality (M) was estimated to be approximately 0.12. Yield-per-recruit analyses indicated that mulloway in New South Wales are being growth overfished and substantial increases in yield could be achieved by increasing the length at first harvest. Values of the spawning potential ratio were below 0.2 under a range of mortality estimates, suggesting that mulloway are at risk of recruitment overfishing. These results suggest that the spawning stock of mulloway in south-eastern Australia has been depleted and that remedial management action is required to protect this iconic species. 相似文献
15.
16.
对驼背鲈(Chromileptes altivelis)的胚胎发育及仔、稚、幼鱼的形态特征进行了详细的观察与研究,描述了从受精卵到仔、稚、幼鱼各发育期的时间和形态特征变化。结果表明,在水温25-26℃、盐度30的海水中,受精卵历时27 h 25 min完成整个胚胎发育过程,经历从卵裂、囊胚、原肠、神经胚到肌节形成、各器官的逐渐形成、变化、完善等一系列的胚胎发育和变化过程;根据其卵黄囊消长情况、鳞片的覆盖状态、体色发生的不同变化以及第二背鳍和腹鳍的消长,将胚后发育分为仔、稚、幼鱼3个阶段。在水温22-26℃、盐度29-31、DO≥5 mg/L的条件下,2-3 d仔鱼卵黄囊消失,开口摄食;生长发育至31 d,仔鱼已变态进入稚鱼期;培育至57 d,稚鱼完成变态,成为幼鱼。第二背鳍棘和腹鳍棘的生长与收缩等石斑鱼类早期发育的共性生长特征及其体表特性体色变化特征为驼背鲈胚后发育过程中最明显的特征。 相似文献
17.
Cludio C. Maretti Adriana R. Leo Ana Paula Prates Eliane Simes Ricardo B.A. Silva Ktia T. Ribeiro Leonardo Geluda Manoel S. Sampaio Fernanda F.C. Marques Anna C. Lobo Luís H. de Lima Leonardo M. Pacheco Warwick A. Manfrinato Antnio Q. Lezama Matheus T.P. Couto Paula M. Pereira Moara M. Giasson Paulo H.M. Carneiro Aldízio L. de Oliveira Filho Bernardo F.A. Brito Maurício S. Pompeu Guilherme F. Dutra Mara C. Nottingham Giovanna Palazzi Fabiana O. Hessel Andr L. Lima Bruna De Vita S. Santos Rodrigo Medeiros Marcelo M. Oliveira Mauro O. Pires Mnica Assad Mariana G. Pereira Marcelo R. Kinouchi Rosana J. Subir 《水产资源保护:海洋与淡水生态系统》2019,29(Z2):44-70
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18.
Charles C. Mischke Matt J. Griffin Terrence E. Greenway David Wise 《Journal of the World Aquaculture Society》2013,44(2):288-292
Mosquitofish, Gambusia sp., have been spread throughout the world to biologically control mosquitoes. However, the fish has gained a reputation as an invasive species and has been implicated in displacing native aquatic species. Gambusia affinis are native to the southeastern United States and commonly occur in commercial channel catfish, Ictalurus punctatus, production ponds. We investigated effects of mosquitofish presence on zooplankton populations, water quality, disease occurrence, and fish production in experimental ponds. There were no differences between ponds with or without mosquitofish in numbers of calanoid copepods, cyclopoid copepods, total copepods, Bosmina sp., Ceriodaphnia sp., Moina sp., Daphnia sp., or total cladocerans. There were also no differences in copepod and cladoceran sizes. Copepod nauplii were more numerous during the summer months in ponds with mosquitofish. There were no differences in water quality variables (soluble reactive phosphorus, nitrate, nitrite, ammonia, total nitrogen, total phosphorus, pH) or phytoplankton density between ponds stocked with and without mosquitofish. Catfish production and disease occurrence were also similar between ponds with and without mosquitofish. Although mosquitofish may cause problems when stocked outside their native range, there does not appear to be any adverse effects of mosquitofish presence in catfish production ponds. 相似文献
19.
D. K. Rowe 《Fisheries Management and Ecology》1999,6(5):377-386
Adult common bullies, Gobiomorphus cotidianus McDowall, are small (total length = 30–120 mm), benthic fish commonly found in the littoral zones of New Zealand lakes where they are a major prey species for trout and eels. Differences in their relative abundance (CPUE) were determined between 21 shallow North Island lakes. Mean CPUE ranged from 2 to > 1000 fish net− 1 night− 1 and was inversely related to water transparency. The abundance of bullies was not reduced in lakes containing rainbow trout, Oncorhynchus mykiss (Walbaum), but was reduced in all lakes containing self-recruiting populations of eels, Anguilla spp. Mean densities of planktonic larval bullies in the limnetic zone were also inversely related to water transparency. Since low water transparency is related to increased trophic status for these lakes, the abundance of bullies is likely to be related to lake productivity, rather than turbidity. 相似文献
20.
《Journal Of Aquatic Food Product Technology》2013,22(4):71-80
No abstract available for this article. 相似文献