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本研究将100~300 pg含有肌肉特异表达启动子和绿色荧光蛋白(Green fluorescent protein,GFP)基因的重组质粒(smyd1:gfp)显微注射到大菱鲆(Scophthalmus maximus)受精卵动物极细胞中,通过细心培育,成功孵化出鱼苗约120尾。统计分析显示,显微注射后,大菱鲆胚胎存活率为4.8%。利用荧光显微镜观察大菱鲆胚胎及仔鱼,只在注射smyd1:gfp质粒的胚胎及仔鱼的肌肉中发现有绿色荧光。通过进一步PCR扩增检测,在注射的大菱鲆胚胎及仔鱼DNA中扩增出了GFP特异片段,大小约为340 bp。研究表明,本研究成功建立了大菱鲆显微注射技术,可为大菱鲆基因功能研究和遗传育种奠定基础。 相似文献
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半滑舌鳎vasa调控区的克隆、表达载体构建及其驱动活性检测 总被引：1，自引：1，他引：0
为研究半滑舌鳎(Cynoglossus semilaevis)vasa(Csvasa)基因调控区的功能,在已克隆的Csvasa基因编码序列的基础上,采用基因组步移和PCR扩增的方法克隆得到Csvasa调控区,通过生物信息学方法分析vasa基因5′区,并构建了含Csvasa基因调控区的绿色荧光蛋白(GFP)表达载体(p Csvasa-GFP-T),进一步通过显微注射技术初步验证调控区的驱动活性。结果表明,通过基因组步移和PCR扩增获得Csvasa 5′区5 166 bp和3′区1 655 bp,利用在线生物信息学软件对5′区序列进行分析,发现在转录起始点上游26 bp处存在保守的TATA框,以及潜在的转录因子结合点如SRY、Oct-1、Sox-5、CREB、GATA、AP-1、C/EBP、Sp-1、c-Myc、HNF、NKX2-5、V-Myb等。通过显微注射技术,将所构建的p Csvasa-GFP-T表达载体注射于青鳉(Oryzias latipes)受精卵并进行培养观测,发现Csvasa调控区能够驱动GFP在青鳉胚胎内表达,荧光表达率为81%。将有荧光的胚胎培养为成鱼,检测外源基因的整合率为11.5%。这些结果为进一步研究半滑舌鳎原始生殖细胞(PGCs)的标记、追踪和操作研究以及半滑舌鳎的性别控制等奠定了基础。 相似文献
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对花鲈(Lateolabrax japonicas)成鱼腹腔注射促黄体激素释放激素类似物(LHRH-A3)和人体绒毛膜促性腺激素(HCG),利用放射性免疫激素测定方法(RIA)检测了花鲈血清中类胰岛素生长因子(Insulin-like growth factor 1,IGF-1)在注射后6、12、24、48 h内的变化情况,同时还利用荧光实时定量PCR技术检测了花鲈肝脏中IGF-1和类胰岛素生长因子结合蛋白1(Insulin-like growth factor-binding protein 1,IGFBP-1)mRNA的相对表达量的变化情况。结果表明,注射LHRH-A3的花鲈血清中IGF-1的含量在24 h后出现了明显的降低(P0.05),而花鲈肝脏中IGF-1 mRNA的相对表达量在注射6 h后较对照组明显升高(P0.05),而肝脏中IGFBP-1 mRNA的相对表达量则呈现下降趋势。注射HCG的花鲈血清中IGF-1的含量在注射后12 h出现了明显降低(P0.05),24 h后血清中IGF-1含量持续降低。肝脏中IGF-1 mRNA的相对表达量在注射后的24 h内均保持稳定,而在处理后的48 h出现了显著上升(P0.05)。肝脏中IGFBP-1 mRNA的相对表达量在注射后12h检测到显著升高(P0.05)。研究结果表明,LHRH-A3和HCG均可以通过直接或间接的刺激作用来触发花鲈IGF系统的应答反应,但是其中具体机制尚不明了。 相似文献
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《渔业科学进展》2014,(4)
对花鲈(Lateolabrax japonicas)成鱼腹腔注射促黄体激素释放激素类似物(LHRH-A3)和人体绒毛膜促性腺激素(HCG),利用放射性免疫激素测定方法(RIA)检测了花鲈血清中类胰岛素生长因子(Insulin-like growth factor 1,IGF-1)在注射后6、12、24、48 h内的变化情况,同时还利用荧光实时定量PCR技术检测了花鲈肝脏中IGF-1和类胰岛素生长因子结合蛋白1(Insulin-like growth factor-binding protein 1,IGFBP-1)mRNA的相对表达量的变化情况。结果表明,注射LHRH-A3的花鲈血清中IGF-1的含量在24 h后出现了明显的降低(P0.05),而花鲈肝脏中IGF-1 mRNA的相对表达量在注射6 h后较对照组明显升高(P0.05),而肝脏中IGFBP-1 mRNA的相对表达量则呈现下降趋势。注射HCG的花鲈血清中IGF-1的含量在注射后12 h出现了明显降低(P0.05),24 h后血清中IGF-1含量持续降低。肝脏中IGF-1 mRNA的相对表达量在注射后的24 h内均保持稳定,而在处理后的48 h出现了显著上升(P0.05)。肝脏中IGFBP-1 mRNA的相对表达量在注射后12h检测到显著升高(P0.05)。研究结果表明,LHRH-A3和HCG均可以通过直接或间接的刺激作用来触发花鲈IGF系统的应答反应,但是其中具体机制尚不明了。 相似文献
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为探究花鲈(Lateolabrax maculatus)干扰素刺激基因 15 (Interferon-stimulated gene 15, ISG15)在抵抗病毒过程的免疫应答反应, 本研究克隆、鉴定了花鲈 ISG15 基因(LmISG15), 并对其进行生物信息学分析, 获得了 LmISG15 在各组织中的表达特征及其在受到传染性脾肾坏死病毒(infectious spleen and kidney necrosis virus, ISKNV)感染后的时空表达特征。结果显示, 该基因全长 1625 bp, ORF 为 480 bp, 编码 160 个氨基酸, 分子质量为 17.5 kD, 理论等电点为 9.41。LmISG15 的氨基酸序列较为保守, 与眼斑拟石首鱼(Sciaenops ocellatus)和条石鲷(Oplegnathus fasciatus)相似度分别为 74.07%和 71.88%。LmISG15 在花鲈 10 种组织中均有表达, 在心脏中表达量最高, 其次在脑、中肾和鳃中的表达量较高, 在肌肉中表达量最低。注射 ISKNV 后, 花鲈头肾组织中病毒拷贝数升高, 同时 LmISG15 在花鲈肝脏、头肾和鳃中的表达量显著上调(P＜0.05)。研究表明, LmISG15 参与了抵抗病毒刺激的免疫应答过程。 相似文献
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为探究ncc、nkcc基因在花鲈渗透调节中发挥的作用,实验通过全基因组鉴定、多重序列比对、系统进化树构建以及蛋白结构预测对花鲈ncc进行了鉴定及序列分析,利用实时荧光定量PCR (qRT-PCR)检测ncc和nkcc在海水、淡水花鲈鳃组织中的表达水平,利用原位杂交技术确定ncc2和nkcc1a在海水及淡水花鲈鳃中的表达位置。结果显示,从花鲈中鉴定出2个ncc基因,即ncc1和ncc2,其编码序列(CDS)长度分别为2 691和3 120bp,编码896和1 039个氨基酸,在进化上具有保守性。ncc2在淡水花鲈鳃组织中的表达量显著高于海水,而nkcc1a在海水花鲈鳃组织中的表达量显著高于淡水,ncc1、nkcc1b、nkcc2在海淡水中的表达量则无显著差异。淡水适应过程中花鲈鳃组织中的ncc2的表达量逐渐上调,而nkcc1a的表达量逐渐下调;海水适应过程则呈现相反的表达趋势。此外,原位杂交结果显示,ncc2和nkcc1a基因分别位于淡水与海水中鳃组织的相邻鳃小片间的鳃丝上皮。以上结果表明,ncc2和nkcc1a基因分别编码淡水及海水花鲈鳃中重要的Na+及Cl 相似文献
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为获得快速生长的泥鳅(Misgurnus anguillicaudatus),研究了泥鳅β-actin基因启动子介导的革胡子鲶(Clarias gariepinus)生长激素(growth hormone,GH)重组基因。采用PCR和RT-PCR技术克隆泥鳅β-actin基因近端启动子、3′-UTR及革胡子鲶GH基因编码区,构建一个长为2 418bp的GH重组基因DPRK。首先,构建了以增强型绿色荧光蛋白(EGFP)基因作为报告基因的重组表达载体p AF(pβ-actin promoter-EGFP),然后转染DF1真核细胞,同时经酶切线性化后显微注射到斑马鱼(Danio rerio)和泥鳅的受精卵中,以评价泥鳅β-actin启动子的启动活性。其次,将重组基因DPRK显微注射到泥鳅受精卵中。结果显示:泥鳅β-actin启动子能在DF1细胞、斑马鱼和泥鳅的受精卵中启动绿色荧光蛋白的表达;泥鳅的荧光表达率(78.44%)显著高于斑马鱼(29.62%);泥鳅受精卵显微注射DPRK 20d后,在m RNA水平检测到GH基因的表达。这表明泥鳅β-actin基因近端启动子具有显著的启动活性,且重组基因DPRK能在泥鳅体内表达,为下一步泥鳅的基因工程育种奠定了理论基础。 相似文献
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Association between the interannual variation in the oceanic environment and catch rates of bigeye tuna (Thunnus obesus) in the Atlantic Ocean 
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下载免费PDF全文 The environmental processes associated with variability in the catch rates of bigeye tuna in the Atlantic Ocean are largely unexplored. This study used generalized additive models (GAMs) fitted to Taiwanese longline fishery data from 1990 to 2009 and investigated the association between environmental variables and catch rates to identify the processes influencing bigeye tuna distribution in the Atlantic Ocean. The present findings reveal that the year (temporal factor), latitude and longitude (spatial factors), and major regular longline target species of albacore catches are significant for the standardization of bigeye tuna catch rates in the Atlantic Ocean. The standardized catch rates and distribution of bigeye tuna were found to be related to environmental and climatic variation. The model selection processes showed that the selected GAMs explained 70% of the cumulative deviance in the entire Atlantic Ocean. Regarding environmental factors, the depth of the 20 degree isotherm (D20) substantially contributed to the explained deviance; other important factors were sea surface temperature (SST) and sea surface height deviation (SSHD). The potential fishing grounds were observed with SSTs of 22–28°C, a D20 shallower than 150 m and negative SSHDs in the Atlantic Ocean. The higher predicted catch rates were increased in the positive northern tropical Atlantic and negative North Atlantic Oscillation events with a higher SST and shallow D20, suggesting that climatic oscillations affect the population abundance and distribution of bigeye tuna. 相似文献
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Liping Li Rui Wang Wanwen Liang Xi Gan Ting Huang Yan Huang Jian Li Yunliang Shi Honglin Luo Ming Chen 《广西水产科技》2014,(1)
Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China. 相似文献
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Iciar Martinez Amaya Velasco Ricardo Pérez-Martín Eskil Forås Michiaki Yamashita Carmen G. Sotelo 《Journal Of Aquatic Food Product Technology》2013,22(4):289-297
The purpose of this work was to evaluate the suitability of a method based on the presence of two restriction sites (for Hae III and Hindf I) in the mitochondrial NADH dehydrogenase subunit 5 (mt ND5) gene to identify Scomber species. The evaluation was performed on 144 reference and market samples by sequencing of the entire 505-bp fragment of the mt ND5 gene and of a 464-bp fragment of the Kocher fragment of the cytochrome b gene (mt Cytb). Sequence analysis of any of the two fragments allows the identification of each of the four Scomber species, but S. japonicus and S. colias had the same restriction sites at the ND5 amplicon and would not have been differentiated by this analysis. Similarly, loss of the Hae III site in some S. scombrus individuals would have misidentified them as not being Scomber. All the market products were correctly labeled except one acquired in Spain labeled as originating in the Atlantic and containing S. japonicus. 相似文献
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In this experiment, a feeding trial was performed to determine the effects of fructooligosaccharide (FOS) on growth performance, digestive enzyme activity and immune response of Japanese sea bass, Lateolabrax japonicus juveniles (initial weight 38.3 ± 0.5 g), and the fish were examined following feeding with six levels of FOS (0, 0.5, 1, 2, 4 and 6 g/kg) for 28 days. Significant enhancement of weight gain (WG) and specific growth rate (SGR) was found in fish fed 1 g/kg FOS incorporated diets (p < .05), while the feed conversion ratio (FCR) in the 1, 2 g/kg FOS groups reduced significantly compared with the control (p < .05). Besides, the crude lipid in the 4, 6 g/kg FOS groups increased significantly compared with the control (p < .05). On the other hand, the erepsin and lipase activities significantly elevated in intestine of fish fed 2 g/kg FOS (p < .05) and the lysozyme activity in serum of fish fed 2 g/kg FOS were significantly higher than that in the control (p < .05). Moreover, the alkaline phosphatase activities in serum of fish fed 0.5, 1, 2 g/kg FOS were significantly higher than in control (p < .05). Regression analysis showed that the relationships between dietary FOS levels and either SGR, FCR, erepsin or lysozyme activities were best expressed by regression equations, and the optimal inclusion levels are 1.37, 1.80, 3.06, 3.11, 1.93 and 1.80 g/kg for SGR, FCR, erepsin, lipase, lysozyme and total superoxide dismutase activities, respectively. Overall, this study revealed that FOS incorporated diets could beneficial for L. japonicus culture in terms of increasing the growth, digestion and immune activities. Under the present experimental condition, the optimal supplementary level of FOS in the diet of L. japonicus is 1–3 g/kg. 相似文献
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Growth hormone (GH) and reproduction: a review 总被引：5，自引：0，他引：5
Florence Le Gac Odile Blaise Alex Fostier Pierre-Yves Le Bail Maurice Loir Brigitte Mourot Claudine Weil 《Fish physiology and biochemistry》1993,11(1-6):219-232
Interaction between growth and reproduction occurs in many vertebrates and is particularly obvious at certain stages of the life cycle in fish. Endocrine interactions between the gonadotropic axis and the somatotropic axis are described, the potential role of GH being emphasised. A comparative analysis of these phenomena in mammals, amphibians and fish, suggests a specific role of GH in the physiology of puberty, gametogenesis and fertility. It also shows the original contribution made by studies on the fish model in this field of investigations. 相似文献
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Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL?1 (1.25 ng well?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation. 相似文献
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Qin-Cheng Huang Di‐Gen Qin Bei‐Ping Tan Tao Du Yuan-Zhi Yang Qi-hui Yang Shu‐Yan Chi Xiao-hui Dong 《Aquaculture Research》2020,51(2):816-827
A 5 × 3 factorial growth trial was conducted to evaluate optimal dietary protein and lipid levels (dietary protein level, DP; dietary lipid level, DL) for juvenile Sillago sihama (S. sihama) (2.0 ± 0.02 g, initial weight). Fish were fed 15 diets containing 5 DPs (350, 400, 450, 500 and 550 g/kg) and 3 DLs (60, 90 and 120 g/kg) for 8 weeks. The interaction between proteins and lipids significantly influenced the feed conversion ratio, condition factor, body composition, antioxidant indices and lipase activity (p < .05). DP 450 g/kg showed the highest average final body weight. DPs 500 and 550 g/kg significantly decreased the protein efficiency ratio (p < .05). DL 120 g/kg showed the highest percentage weight gain. The low feed conversion ratio was found in diets P45L12, P55L9 and P55L12. Diet P45L12 showed high superoxide dismutase activities. DP 450 g/kg showed the lowest average malondialdehyde content. Lipase activity was increased by increasing DP (p < .05) with a fall at DP 550 g/kg. Under the present experimental conditions, the optimal DP for S. sihama was 450 g/kg under the DL 120 g/kg. 相似文献