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Aliza D Ismail IS Kuah MK Shu-Chien AC Tengku Muhammad TS 《Fish physiology and biochemistry》2008,34(2):129-138
Copper is one of the major heavy metal pollutants found in the aquatic environment. Therefore, it is important for determining
the genes that play a key role in copper metabolism in aquatic organisms. This study, thus, aimed to identify a new copper-inducible
gene in swordtail fish, Xiphophorus helleri. Using ACP-based RT-PCR coupled with RLM-RACE, we cloned Wap65, a mammalian homologue of hemopexin gene. The gene exhibits
high identity at amino acid levels with the Wap65 gene of other fish species (42–68%) and mammalian hemopexin gene (35–37%).
In addition, ten cysteine and two histidine residues are conserved in the swordtail fish Wap65 gene. These cysteine residues
are vital for structural integrity, and histidine residues provide high binding affinity towards heme. As revealed by RT-PCR,
the gene was upregulated in swordtail fish that were exposed to copper in a dose- and time-dependent manner. Therefore, the
identification of Wap65, a mammalian homologue of hemopexin, as a new copper-inducible gene will provide greater insight into
the role of this gene in copper metabolism. 相似文献
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肌原纤维结合型丝氨酸蛋白酶(myofibril-bound serine proteinase,MBSP)是最近发现的一种蛋白酶。该酶参与肌原纤维蛋白的降解及鱼糜制品弹性的下降。但是,对该酶一级结构的研究,迄今为止,未有报道。本文根据已测定的鲤MBSP N-末端氨基酸序列以及丝氨酸蛋白酶活性中心保守序列设计兼并引物,结合RT-PCR技术实现了MBSP基因片段的扩增。再根据克隆到的MBSP片段序列设计基因特异引物,用于MBSP基因的5′和3′末端快速扩增。综合以上结果,鲤MBSP的全长被确定。序列分析表明,MBSP cDNA含有一732 bp的开放阅读框,编码243个氨基酸残基,其中信号肽长度为21个氨基酸残基。组成丝氨酸蛋白酶活性中心的氨基酸残基(His61,Asp107和Ser197)在MBSP中保守存在。成熟MBSP含有222个氨基酸残基,分子量为24.5 ku,比其天然蛋白的分子量30 ku略小。成熟MBSP的等电点为10.43。鲤MBSP与鲫MBSP,猪胰蛋白酶,牛胰蛋白酶,美洲鲽胰蛋白酶的同源性分别为80.6%,55.8%,55.3%和53.9%。而与仓鼠肌肉中具有胰凝乳蛋白酶性质的蛋白酶的同源性为39.2%。MBSP有高含量的赖氨酸残基(11.93%),此特性可能与该酶的肌原纤维结合特性有关。 相似文献
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Tomoya Kono Yoichi Kitao Kohei Sonoda Royhei Nomoto Tohru Mekata Masahiro Sakai 《Fisheries Science》2008,74(3):603-612
ABSTRACT: A ghrelin gene has been cloned and sequenced in common carp Cyprinus carpio . Ghrelin cDNA is composed of 461 bp [with a 36-bp 5'-untranslated region (UTR) and a 113-bp 3'-UTR], which translates into a protein of 103 amino acid residues. Carp ghrelin (preproghrelin) contained a predicted signal peptide of 26 amino acid residues, the ghrelin domain ( Gly 27 – Val 45 ) and C-terminal peptide ( Gly 46 – Phe 103 ). Homology analysis of the ghrelin domain of carp with that of other known ghrelin in vertebrates showed good similarity to teleost ghrelin (50–81.8%). Hydropathy analysis based on the deduced amino acid sequence of ghrelin domains in teleosts showed a similar profile. Carp ghrelin clustered with ghrelin of goldfish Carassius auratus and other teleosts, away from mammalian, reptilian, avian, amphibian and chondrichthian ghrelin, by phylogenetic analysis. Genomic organization of carp ghrelin gene was composed of four exons and three introns, which was the same as that of other teleosts and human ghrelin genes. The carp ghrelin gene was expressed in unstimulated tissues such as foregut, hindgut, spleen and brain. In spleen cells, expression of the ghrelin gene increased upon stimulation with lipopolysaccharide (LPS), phytohemagglutinin (PHA) or imiquimod. The identification of carp ghrelin gene and the analysis of the modulation of its expression in immune-activated conditions will allow a more complete analysis of the roles of ghrelin in teleosts. 相似文献
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Effects of temperature and dietary protein on gene expression of Hsp70 and Wap65 and immunity of juvenile mirror carp (Cyprinus carpio) 
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下载免费PDF全文 Juvenile mirror carp were fed diets containing 303.4, 321.7, 341.2, 361.0 and 379.1 g kg?1 proteins, respectively, and reared at different water temperatures (18, 23 and 28°C) for 60 days. Gene expression of heat shock protein gene (Hsp70) and the warm temperature acclimation‐related 65 kDa protein gene (Wap65), immunity and antioxidant status in the carp were investigated. Results indicated that the contents of serum complement 3 (C3), complement 4 (C4) and immunoglobulin M (IgM), as well as activities of liver superoxide dismutase (SOD) and lysozyme (LSZ) were significantly enhanced with increasing dietary protein (P < 0.05), while content of malondiadehyde (MDA) decreased. Gene expression level of Wap65 in the liver significantly increased with dietary protein, while gene expression of Hsp70 decreased. The contents of C3, C4 and IgM, the activities of SOD and LSZ and gene expression level of Wap65 in the liver significantly increased with temperature. These results suggest that: Serum immune parameter, antioxidant enzymes and Hsp70 and Wap65 expression interact in fish to improve ability to adapt to the environment; and the optimal conditions for the immunity of carp are 348.1?354.5 g kg?1 protein at 18°C, 352.3?364.9 g kg?1 at 23°C and 360.2?364.3 g kg?1 at 28°C, and the optimum temperature for carp is 23°C. 相似文献
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Erika Cottone Valentina Pomatto Fulvio Cerri Ezio Campantico Ken Mackie Massimiliano Delpero Alda Guastalla Claudio Dati Patrizia Bovolin Maria Fosca Franzoni 《Fish physiology and biochemistry》2013,39(5):1287-1296
Cannabinoids, the bioactive constituents of Cannabis sativa, and endocannabinoids, among which the most important are anandamide and 2-arachidonoylglycerol, control various biological processes by binding to specific G protein-coupled receptors, namely CB1 and CB2 cannabinoid receptors. While a vast amount of information on the mammalian endocannabinoid system does exist, few data have been reported on bony fish. In the goldfish, Carassius auratus, the CB1 receptor has been cloned and its distribution has been analyzed in the retina, brain and gonads, while CB2 had not yet been isolated. In the present paper, we cloned the goldfish CB2 receptor and show that it presents a quite high degree of amino acid identity with zebrafish Danio rerio CB2A and CB2B receptors, while the percentage of identity is lower with the puffer fish Fugu rubripes CB2, as also confirmed by the phylogenetic analysis. The sequence identity becomes much lower when comparing the goldfish and the mammalian CB2 sequences; as for other species, goldfish CB2 and CB1 amino acid sequences share moderate levels of identity. Western-blotting analysis shows the CB2 receptor as two major bands of about 53 and 40 kDa and other faint bands with apparent molecular masses around 70, 57 and 55 kDa. Since the distribution of a receptor could give information on its physiological role, we evaluated and compared CB1 and CB2 mRNA expression in different goldfish organs by means of qReal-Time PCR. Our results show that both CB1 and CB2 receptors are widely expressed in the goldfish, displaying some tissue specificities, thus opening the way for further functional studies on bony fish and other nonmammalian vertebrates. 相似文献
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The objective of this study was to determine the growth of Trypanosoma danilewskyi (Laveran & Mesnil, 1904) from goldfish, Carassius auratus (L.), in medium containing no serum, or in medium supplemented with either 10% fish serum (goldfish, carp, or tin foil barb) or 10% mammalian serum (horse or foetal bovine). After 10 days, the number of trypanosomes in flasks containing tin foil barb serum increased by nearly 700% over the initial inoculum. Similarly, a substantial increase in the number of parasites was seen after 10 days in the cultures containing carp and goldfish serum. After 6 weeks, there was more than a 15-fold increase of T. danilewskyi in cultures containing goldfish serum. Medium containing no serum or mammalian serum failed to support the growth of parasites. Therefore, serum-related factors within fish blood are required for the propagation of T. danilewskyi isolated from infected goldfish. Since T. danilewskyi can be propagated in vivo and in vitro in the presence of homologous proteins, cultured and wild-type forms can be compared to determine if cultured parasites can be used as analogues of natural life-cycle stages. 相似文献
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生长抑素具有抑制脑垂体GH释放的作用，是调控鱼类生长的主要激素之一。本研究采用RT和RACE(rapid amplification of cDNA ends)法，分离和测定了团头鲂脑中生长抑素PSSI cDNA的全长核苷酸序列，并对该基因进行了结构和系统进化分析。3’RACE扩增得到700bp左右的片段，5’RACE分离得到500bp左右的片段，把3’片段与5’片段拼接得到全长cDNA。cDNA全长735bp[不含poly(A)]，5’端非翻译区有100nt。3’端290bp[不包含poly(A)]，阅读框(open reading frame，ORF)345bp。该序列与金鱼PSSI cDNA序列同源性为90％，主要差异在5’端非翻译区。团头鲂生长抑素mRNA阅读框编码114个氨基酸，包括一些酶切位点，产生26个氨基酸的大分子态生长抑素，进一步加工成与人等结构相似的14个氨基酸的生长抑素；团头鲂生长抑素前体氨基酸序列与金鱼、虹鳟、鲶鱼、鲅鱇、蛙、牛、鼠、鸡、猴、人等的比较发现，它与金鱼的同源性最高达95％，与鮟鱇最低50％，与人68％。这说明生长抑素基因在长期的进化中相当保守，同时，也因为鱼类生活环境多样导致基因变异较大。 相似文献
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为全面了解密云水库鲢鱼、鳙鱼的营养特征,以密云水库秋季鲢鱼和鳙鱼为研究对象,对其肌肉的水分、蛋白质、粗脂肪、灰分、矿物质元素含量以及氨基酸、脂肪酸组成进行分析。研究结果表明,密云水库鲢鱼和鳙鱼肌肉蛋白质含量分别为(18.40±0.40)%和(18.60±0.57)%,粗脂肪含量为(0.60±0.10)%和(1.48±0.91)%,富含人体所需的钾、钠、钙、镁、磷等常量元素及铁、锰、锌、硒等微量元素;鲢鱼、鳙鱼肌肉中,18种常见氨基酸均被检出,其中谷氨酸含量高,必需氨基酸分别占氨基酸总量的40.00%和40.16%,必需氨基酸指数分别为99.15和98.76;鲢鱼、鳙鱼肌肉中分别检出脂肪酸12种和19种,相对含量均表现为多不饱和脂肪酸>饱和脂肪酸>单不饱和脂肪酸,而多不饱和脂肪酸中均以二十二碳六烯酸相对含量最高,其次为二十碳五烯酸,两者之和分别达30.64%和23.79%。研究结果表明,密云水库鲢鱼、鳙鱼是良好的蛋白质来源,氨基酸组成合理,富含矿物质和有益脂肪酸,具有很好的营养价值和可食用性。 相似文献
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一种贝类作为水产动物诱食剂的研究 总被引：7，自引：0，他引：7
一种贝类（ＡＳＬ）不仅蛋白质含量高,氨基酸组成也比较全面,高不饱和脂肪酸含量丰富,甜菜碱含量高达２８９ｍｇ／ｇ。向饲料中添加１％的ＡＳＬ粉,发现其对金鱼的诱食活性不仅显著高于空白对照组,而且显著高于化十香组;饲料中添加０３％的ＡＳＬ粉,异育银鲫摄食量显著高于对照组,六周增重率比对照组提高３３％,这些结果表明ＡＳＬ作为水产动物诱食剂具有很高的开发利用价值。 相似文献
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Raffaella Branciari Eleonora Urbani Andrea Valiani Massimo Trabalza-Marinucci Alessandro Dal Bosco 《Journal Of Aquatic Food Product Technology》2017,26(6):686-694
The nutritional profile and the consumer preference of fish burgers made from carp, goldfish, perch, and tench were investigated. Perch and goldfish burgers, which have the highest proportion of eicosapentaenoic acid (6.63% and 4.90%, respectively) and docosahexaenoic acid (20.28% and 11.99%, respectively), showed lower absolute amounts of these fatty acids (17.03 and 10.10 mg/100 g food, respectively) than tench and carp (31.16 and 49.63 mg/100 g food, respectively). The consumer evaluation established that differences in the perception of the fish burgers exist. The overall liking and taste were the main attributes that influenced both consumer’s acceptance and purchase intent. 相似文献
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S. L. Hsieh H. T. Chang C. H. Wu C. M. Kuo 《Aquaculture (Amsterdam, Netherlands)》2004,230(1-4):527-546
The stearoyl-CoA desaturase cDNA in tilapia (Oreochromis mossambicus) was cloned by RT-PCR and RACE, and it was compared with those in grass carp, common carp and milkfish. Nucleotide sequence analysis revealed that the full length of cDNA (1172 bp) clone encompasses 1008 bp open reading frame (ORF) encoding 336 amino acid residues. The deduced amino acid sequence shares 78–82% identity with the teleosts and 64–66% with mammals compared, and like these fish, the cloned tilapia stearoyl-CoA desaturase amino acid sequence conserves three histidine cluster motifs (one HXXXXH and two HXXHH), which functioned as non-heme iron binding sites, essential for stearoyl-CoA desaturase activity. RT-PCR and Northern blot analysis reveal that tilapia stearoyl-CoA desaturase is expressed only in liver, but the stearoyl-CoA desaturase expression in multiple tissues was observed in milkfish, grass carp and carp. Further, the hormonal regulation of stearoyl-CoA desaturase gene expression was investigated by a single injection of 17β-estradiol and testosterone. The results showed that the administration of 17β-estradiol to tilapia led to a greater increase in desaturase activity than testosterone, and higher doses of steroids produced greater increases in enzyme activity. The comparative RT-PCR analysis showed that the stearoyl-CoA desaturase mRNA level increased significantly in 17β-estradiol treated animals, especially in the groups receiving a single injection of 50 mg 17β-estradiol. This was reflected in the decrease in the saturated fatty acids and the increase in the monounsaturated fatty acids. The proportion of the polyunsaturated fatty acids was not affected. 相似文献
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Whole-body amino acid composition and comparative aspects of amino acid nutrition of the goldfish, golden shiner and fathead minnow 总被引：2，自引：0，他引：2
Delbert M. Gatlin III 《Aquaculture (Amsterdam, Netherlands)》1987,60(3-4):223-229
Pond-raised fingerling goldfish (Carassius auratus), golden shiners (Notemigonus crysoleucas) and fathead minnows (Pimephales promelas) were analyzed for whole-body amino acid composition. All three species had similar amino acid patterns in whole-body tissue. These compositional data were also used to compute essential amino acid ratios (A/E ratios) which were compared with the quantified amino acid requirements of another cyprinid species, the common carp (Cyprinus carpio). The whole-body A/E ratios of the three baitfish species were very similar to the A/E ratios of the carp requirement values. Since previous studies have established a high correlation between whole-body amino acid patterns and dietary amino acid requirement patterns of several fish species, the whole-body amino acid values determined in this study will provide indices for formulating diets for the goldfish, golden shiner and fathead minnow. 相似文献
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根据已经获得的两种鲢肌球蛋白重链同工型基因(低温型sc-w和高温型sc-s)在3′端展现的明显差异,设计了2个特异性的反向引物,以鲤科鱼类肌球蛋白重链5′端的保守序列为正向引物,通过Long-PCR对编码鲢两种肌球蛋白重链同工型的球状结构域(Subfragment-1,S1)的全长基因进行了克隆和测序,并推断出它们一级结构的氨基酸序列。研究结果表明,sc-w与sc-s在S1的初级结构上显示80.5%的同源性、与已经报道的草鱼低温型(gc10)有97.2%的高同源性;sc-s则与草鱼中间型(gcI)和高温型(gc30)显示了分别为98.4%和97.1%的高同源性。低温型的sc-w和gc10在S1初级结构上展现的特有变异主要发生在43个氨基酸残基位点,其中15个属保守性残基。对S1区域中两个功能性的表面环loop1(与ATP结合位点有关)和loop2(与肌动蛋白结合位点有关)的结构解析发现,sc-w和gc10在两个表面环的长度、残基电荷分布和氨基酸组成等方面与其它同工型之间存在明显差异,揭示了这两个表面环的结构差异可能影响了栖息于不同环境温度下的淡水鱼的肌球蛋白分子马达功能。分子系统树的分析结果进一步证明,鱼类栖... 相似文献
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夏花青鱼饵料中的最适蛋白质含量 总被引：15，自引：0，他引：15
用酪蛋白、结晶氨基酸、鱼肝油、糊精、纤维素粉等加上适量的维生素、矿物混合剂所组成的精制饵料，以青鱼夏花为试验对象，经过八个星期的喂养试验，从生长、增重和饵料利用率等指标来评定，青鱼夏花的最适蛋白质含量应为41％。蛋白质含量太高，反而减慢生长，降低成活率，同时似乎还对鱼类有毒害影响。蛋白质含量较低，虽然能很好地利用蛋白质，但鱼生长缓慢。6％以下的蛋白质含量甚至出现减重现象。根据本次试验结果，可以认为二龄青鱼和老口青鱼应用饵料中蛋白质的含量为33％和28％是适当的。 相似文献
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用两种粪便收集方法测定草鱼对九种饲料原料的表观消化率 总被引：3，自引：0，他引：3
在(21.5±1.0)℃水温下,以Cr2O3为指示物采用自排法和挤压法分别收集平均体重为65 g草鱼的粪便,测定草鱼对棉籽粕、花生粕、双低菜籽粕、秘鲁鱼粉、菜籽粕、小麦次粉、米糠、大豆粕和肉骨粉9种饲料原料主要营养成分的表观消化率。试验饲料以70%基础饲料和30%的待测饲料原料组成。结果表明:(1)草鱼对肉骨粉、秘鲁鱼粉、棉籽粕、大豆粕的干物质表观消化率较高,分别为60.69%、59.07%、61.08%、57.99%;(2)用自排法和挤压法测定草鱼对饲料原料的表观消化率时,干物质表观消化率有差异,而饲料原料粗蛋白质表观消化率基本保持一致;(3)草鱼对各种饲料原料的蛋白质表观消化率与氨基酸表观消化率的趋势基本一致。 相似文献
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池塘和稻田养殖模式对金边鲤和建鲤肌肉品质的影响 总被引：3，自引：0，他引：3
分析了稻田和池塘养殖模式下金边鲤和建鲤肌肉营养成分、理化特性和质构特性,并进行营养价值评价,评估金边鲤的肌肉品质,为开发和推广金边鲤稻田养殖提供数据依据。结果显示,稻田金边鲤的粗脂肪、粗灰分和系水力均显著高于池塘金边鲤;稻田建鲤和池塘建鲤的基本营养成分除水分外差异均不显著;稻田金边鲤肌肉的硬度、弹性、咀嚼性显著高于池塘建鲤和稻田建鲤,硬度和恢复性显著高于池塘金边鲤,而内聚性显著低于池塘金边鲤、池塘建鲤和稻田建鲤;测定的17种氨基酸中,4组鱼肉的Lys含量均高于WHO/FAO标准和鸡蛋蛋白标准,稻田金边鲤的∑TAA、∑EAA、∑DAA和∑NEAA均高于池塘建鲤和稻田建鲤,且EAA/TAA比值更接近FAO/WHO要求的40%的标准;稻田和池塘金边鲤的EAAI均超过100分,高于稻田和池塘建鲤(82.31～83.36分);测定出的24种脂肪酸中,稻田金边鲤的∑SFA、∑MUFA和DHA+LA+油酸总量均高于其他3组。研究表明,金边鲤和建鲤鱼肉可作为人体优质的赖氨酸源,稻田金边鲤肌肉比建鲤和池塘金边鲤肌肉更富有嚼劲,其肌肉必需氨基酸、不饱和脂肪酸及高度不饱和脂肪酸含量更高,且组成比例更符合人体需求。稻田养殖金边鲤的肌肉品质优势明显,具有较高的产业开发潜力。 相似文献