共查询到20条相似文献，搜索用时 531 毫秒
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Song-Lin Chen Mei-Yu Xu Song-Nian Hu Lin Li 《Aquaculture (Amsterdam, Netherlands)》2004,240(1-4):115-130
Expressed sequence tag (EST) analysis is an efficient tool for gene discovery and for profiling gene expression. In order to isolate functional genes involved in immunity in fish, a cDNA library was constructed from red sea bream (Chrysophrys major) spleen by unidirectional cloning. A total of 2010 ESTs from the library was sequenced and compared with sequences in the GenBank database. Of the 2010 ESTs, 320 ESTs (15.9%) were identified as orthologs of known gene from other organisms by BLAST searches, whereas 1690 ESTs (84.1%) appeared to be unknown and are likely to represent newly described genes. These identified clones were derived from at least 81 genes, which were categorized into eight categories: 9 in cell structure/motility (11.1%), 14 in metabolism (17.3%), 8 in cell defense/immunity (10%), 5 in cell division (6.2%), 7 in cell signal transduction/communication (8.6%), 30 in gene/protein expression (37%), 5 hemoglobin (6.2%) and 3 genes lacking enough information to be classified (3.7%). Several important cDNAs involved in immune functions, such as immunoglobulin light chain (IgL), MHC class II, MHC class IIβ and RAP2c, were identified in red sea bream and compared for their structure with those from other organisms. Alignment showed that the red sea bream IgL precursor was closer to that of spotted wolfish than to that of yellowtail, Europe sea bass, orange spotted grouper, Atlantic salmon, channel catfish, fugu and sterlet. Phylogenetic analysis indicated that the red sea bream MHC II and MHC IIβ were more related to those from striped sea bass than to those from cichlid, flounder, salmonids, zebrafish and carp. High identity (over 92%) in deduced amino acid sequence of RAP2c between red sea bream and mammals implied that RAP2c gene was highly conserved during evolution. 相似文献
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ABSTRACT: Fish transgenesis has progressed considerably. However, the technique of gene transfer for most marine aquaculture species has not been established. A method to introduce foreign genes into fertilized eggs of Japanese flounder Paralichthys olivaceus by particle gun bombardment was developed by the authors. A recombinant plasmid which contains the Japanese flounder keratin gene promoter linked to the green fluorescence protein (GFP) gene was introduced into fertilized eggs by particle gun bombardments twice at 250 psi. In each experimental group, 25 000–35 000 eggs were treated. However, the survival rate was 12.8% which is lower than that of the control groups (56.8%). Of 2606–5205 the embryos survived for 24 h, 43–61 GFP positive embryos were obtained in one experiment, giving a final gene transfer efficiency of 1.4%. All GFP-positive embryos developed and hatched normally. GFP expression was observed in epithelial tissue throughout the developmental stages. At 3 months after gene transfer, foreign DNA was detected by genome polymerase chain reaction analysis in 37 of 69 fry (53.6%). These results suggest that the particle gun method is an effective method to use with fertilized Japanese flounder eggs. 相似文献
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ABSTRACT: Interleukin-1β (IL-1β) cDNA of Japanese flounder was found to consist of 1329 bp, encoded 247 amino acid residues. Among the fish IL-1β in the databases, the one with the highest identity of Japanese flounder IL-1β was that of seabass (62% identity). The expression of IL-1β was induced by treatment with concanavalin A (ConA)/phorbol myristate acetate (PMA) and lipopolysaccharide. The copy number of IL-1β mRNA was increased 30-fold after stimulation with ConA/PMA. Of 871 cDNA on a microarray, 93 genes (10.7%) were up-regulated or down-regulated by IL-1β at 1, 3 and 7 days post-injection. The induced gene expression was highest on day 1 followed by day 3 and day 7. A total of 7% of known and 3.7% of unknown genes of the 871 tested genes were differentially expressed. Of the genes tested, 7.4% were up-regulated and 3.3% were down-regulated. Cytokine genes such as tumor necrosis factor, granulocyte colony stimulating factor and chemokine receptor A were induced in response to IL-1β. Cell surface antigens such as IgM, MHC class I and CD20 receptor were up-regulated. Signal transduction genes such as Toll-like receptor 1 and SH3P2 were also up-regulated. The glucocorticoid receptor and cAMP early repressor were down-regulated in our microarray analysis. 相似文献
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大菱鲆脾脏cDNA文库构建、EST序列分析与免疫抗病相关基因的筛选 总被引：1，自引：0，他引：1
以大菱鲆脾脏为材料,构建cDNA文库,通过对文库克隆的序列测定和初步生物信息学分析,共获得3 656个大菱鲆脾脏表达序列标签（EST）。经与GenBank数据库的序列比对后发现,1 891个EST代表了149个已知基因,其余1 765个EST为未知序列。这149个基因大致可分为6类：9个为细胞结构类（6.0%）;26个为代谢类（17.5%）;45个为细胞防御/免疫类（30.2%）;43个为基因/蛋白质表达类（28.9%）;16个为细胞信号转导/细胞交流类（10.7%）;10个无法明确分类（6.7%）。鉴定的与免疫抗病相关重要基因主要有：CC/CXC趋化因子;主要组织相容性复合体（MHC）class I α;C3补体;天然杀伤细胞增强因子（NKEF）;胸腺素β-4/β-12;干扰素诱导蛋白Gig1;白介素8受体;STAT 4;热激蛋白70/90等。 相似文献
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cDNA cloning of Japanese oyster stress protein homologous to the mammalian 78-kDa glucose regulated protein and its induction by heatshock 总被引：1，自引：0，他引：1
Yoshihiro Yokoyama Hisashi Hashimoto Satoshi Kubota Akira Kuriyama Yoichi Ogura Shoshi Mizuta Reiji Yoshinaka Haruhiko Toyohara 《Fisheries Science》2006,72(2):402-409
ABSTRACT: The 78-kDa glucose regulated protein (GRP78), a member of stress proteins, was cloned from a cDNA library of Japanese oyster Crassostrea gigas . The analysis on Japanese oyster GRP78 clone of approximately 2.6 kb revealed that the entire open reading frame was 1983 bp long and encoded 661 amino acid residues. At the DNA sequence level, the coding region of Japanese oyster GRP78 gene was 72, 62, and 62% identical to those of chicken GRP78, Japanese flounder HSP70, and Japanese flounder HSC71 genes, respectively. Deduced amino acid sequence of Japanese oyster GRP78 was 84, 62, and 62% identical to those of chicken GRP78, Japanese flounder HSP70, and Japanese flounder HSC71, respectively. Japanese oyster GRP78 contained an 18-residue sequence at the N-terminus that exhibits characteristics of a cleavable signal sequence. It also contained an ATPase domain, and a peptide-binding domain in addition to a Lys-Asp-Glu-Leu (KDEL) peptide motif that is involved in determining endoplasmic reticulum localization. Northern blot analysis showed that GRP78 mRNA was induced with heatshock treatment in the oyster tissues. 相似文献
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Manami Inaba Takeshi Kimura Rika Kikukawa Mitsuko Iwasaki Masato Nose Satoru Suzuki 《Fisheries Science》2007,73(3):615-622
ABSTRACT: Seasonal changes in the infection state of marine birnavirus (MABV) in Japanese flounder Paralichthys olivaceus and in rearing sea water are described. Sea water and 10–11 healthy fish were sampled monthly from April 2002 to February 2003. The MABV genome was detected throughout the year in > 80% of the fish examined at each sampling. The virus was isolated from the liver, kidney, and spleen, but not from the brain. The detection rate in each organ increased from April to October, and then decreased. Detection of virus antigen by the indirect fluorescent antibody technique also showed that the virus was present from spring to autumn (June–September) in the liver, kidney, and spleen, but not the brain. Sequence analysis of the MABV genome at the VP2–NS region revealed two specific mutations compared to the standard yellowtail strain (Y-6). It is suggested that the infection state of MABV in Japanese flounder changes to a latent or persistent infection after autumn. MABV was detected in sea water between September and February, suggesting that virus particles in the environment are relatively higher during cool seasons. 相似文献
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ABSTRACT: The effect of dietary taurine on juvenile Japanese flounder was determined by feeding three taurine-supplemented experimental diets (TAU) and a commercial diet (CD) to evaluate a practical diet for juvenile Japanese flounder. Juvenile Japanese flounder were reared on the three experimental diets supplemented with taurine at 0, 0.5, 1.0% and CD. These diets were fed to juvenile Japanese flounder of an initial mean body weight of 0.2 g for 6 weeks at 20°C and the taurine contents of the whole body and tissues were analyzed. The final average body weight of juvenile Japanese flounder fed the 1.0% TAU was significantly higher than that of the other groups. Taurine contents in the whole body and tissues increased with the increase in dietary taurine level. These results indicate that juvenile Japanese flounder require at least 15 mg/g taurine in the diet, even though a combined mix of fish, krill and squid meal was the main protein source in the experimental diets. 相似文献
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ABSTRACT: The acute toxicities of boron were examined for Japanese flounder Paralichthys olivaceus and red sea bream Pagrus major in terms of fish size and water temperature. Japanese flounder of 0.1–70.0 g and red sea bream of 0.6–20.3 g were exposed to different concentrations of boron for 96 h at 20°C under semi-static conditions. In both fish species, the median lethal concentration (LC50 ) for 96 h of boron increased linearly with increasing fish size, ranging from 108 to 252 mg B/L for the flounder, and from 97 to 172 mg B/L for the sea bream. The effect of water temperature on acute toxicity was examined for Japanese flounder of 0.6 and 1.5 g at 10, 15, 20 and 25°C, and for red sea bream of 0.6 and 2.4 g at 12, 15, 20 and 25°C. The toxicity of boron for the flounder increased linearly with increasing water temperature. The 96 h LC50 values ranged from 299 to 108 mg B/L for the 0.6 g flounder and from 350 to 113 mg B/L for the 1.5 g flounder. A similar trend was shown for the 2.4 g red sea bream; however, the relationship for the 0.6 g red sea bream was not significant. 相似文献
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A cost-effective method for producing and using recombinant growth hormone (GH) has been developed. A cell-surface protein
display system was constructed to produce recombinant Japanese flounder growth hormone (r-JfGH) in yeast Saccharomyces cerevisiae and the biological activity of expressed r-JfGH was evaluated by oral delivery to juvenile Japanese flounder. The expression
vector pYD-E2, containing the JfGH gene, S. cerevisiae GAL1 promoter, and the S.cerevisiae URA3 gene downstream and upstream sequences, was designed and constructed to integrate the GH gene into the yeast genome
and secrete r-JfGH stably on the surface of the yeast cell. Expression of r-JfGH was confirmed by flow cytometry until the
50th continuous generation of transgenic yeast strain y-E2, and over 0.98 μg r-JfGH per 1 mg total protein of y-E2 was estimated
by an ELISA–RA assay. Growth of Japanese flounder was promoted significantly by the direct supplementation of freeze-dried
transgenic yeast into the Japanese flounder juvenile feed at levels of 0.5 and 1%. The weight-specific growth rate and feed
conversion ratio of the treated groups were 23.07 and 21.57% (0.5% feeding level) and 28.85 and 41.18% (1% feeding level),
respectively, higher than that of the control group in a 7-week bioactivity study. 相似文献
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ABSTRACT: The nucleotide sequences of Japanese flounder Paralichthys olivaceus , major histocompatibility complex (MHC) cDNA, classical MHC class Iα, non-classical MHC class Iβ, MHC class IIα and IIβ, were determined. The domain structures and antigen binding motifs of vertebrate MHC are conserved in the Japanese flounder MHC. A phylogenetic analysis supports the classification of these genes into class I and class II MHC. Classical MHC class Iα was ubiquitously expressed, whereas the non-classical MHC class Iβ was expressed mainly in lymphoid organs, gills, intestine and stomach. The MHC classes IIα and IIβ were also ubiquitously expressed. 相似文献
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为了研究牙鲆白化发生过程中的分子调控机制,本研究在获得正常和白化牙鲆转录组及micro RNA(mi RNA)深度测序数据的基础上,对tyrosinase related protein 1(tyrp1)和mmu-mi R-143-5p_R+2(mmu-143)进行了表达模式、靶基因预测及验证分析。首先通过RACE方法克隆得到白化相关基因tyrp1的2个转录本,进化树分析表明这2个转录本分别是tyrp1a和tyrp1b,利用RNAhybrid软件预测到mmu-143可能与tyrp1a基因存在互作关系,通过双荧光素酶实验初步验证了这一靶向关系。进一步的荧光定量结果显示,tyrp1a基因在正常牙鲆皮肤中的表达量显著高于白化牙鲆皮肤的表达量,正常牙鲆皮肤中mmu-143的表达量显著低于白化牙鲆皮肤中的表达量。本研究发现,牙鲆tyrp1存在2个转录本,分别是tyrp1a和tyrp1b。双荧光素酶实验和定量PCR分析初步证实,tyrp1a是mmu-143的靶基因,mmu-143是通过调控tyrp1a基因的表达来影响牙鲆白化的。此研究结果为深入揭示牙鲆白化发生的分子机制提供重要的基础资料。 相似文献
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Seven isonitrogenous and isoenergetic experimental diets were formulated to investigate the effect of low molecular weight fish protein hydrolysate (FPH) in diets on growth performance, feed utilization and liver IGF‐I mRNA levels in Japanese flounder (38.80 ± 1.11 g) fed with high plant protein diets. Fish meal protein was, respectively, replaced by 6% (FPH6), 11% (FPH11), 16% (FPH16), 21% (FPH21), 26% (FPH26) FPH of total dietary protein. FPH diets contained a constant high level of plant protein (690 g kg?1) from soybean meal. As a positive control diet, FM2 contained about 590 g kg?1 plant protein and 410 g kg?1 fish meal protein, while negative control diet FM1 contained about 690 g kg?1 plant protein and 310 g kg?1 fish meal protein. The expression levels of liver IGF‐I mRNA were evaluated using real‐time PCR normalized against the 18S rRNA gene. The results showed that moderate low molecular weight FPH (FPH11) improved growth performance and protein retention. Fish fed with FPH11 and control diet FM2 had similar growth and feed utilization, while high‐level low molecular weight FPH did not improve growth performance and protein retention, and depressed liver IGF‐I mRNA expression in Japanese flounder. 相似文献
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Masayuki YAMAMOTO Hiroyasu MAKINO Tetsushi KAGAWA Osamu TOMINAGA 《Fisheries Science》2004,70(6):1089-1097
ABSTRACT: The occurrence and distribution of larval and juvenile Japanese flounder Paralichthys olivaceus and major demersal fish from May to September were examined at seven sandy beaches at the central Seto Inland Sea. The flounder appeared from late May to August and were mainly collected at a depth of 2–5 m. Settlement of larval flounder was seen from late May to late June. A total of 6412 individuals of 17 species representing 12 families were collected in the study area. The dominant fish species were Favonigobius gymnauchen (56.0%), Tarphops oligolepis (8.1%), Rudarius ercodes (7.8%), P. olivaceus (7.3%), Repomucenus spp. (7.0%) and Heteromycteris japonica (3.3%). The catches of flounder differed among beaches and months. The number of flounder in June at Ohama Beach, where the most flounder were collected, was 22.6 individuals per haul (400 m2 ). The density of the flounder correlated negatively to that of F. gymnauchen . 相似文献
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迟缓爱德华氏菌病是海水鲆鲽鱼类的主要病害,发掘抗病分子标记辅助育种是十分有效的策略。本研究以牙鲆(Paralichthys olivaceus) rnd1基因(Pornd1)为对象,对该基因在牙鲆抗病免疫方面的作用进行系统分析。首先对Pornd1基因进行克隆鉴定和抗病相关单核苷酸多态性位点的定位,然后利用荧光定量PCR (qRT-PCR)对Pornd1基因的组织分布、细菌感染后表达情况以及在抗病和易感家系中的表达水平进行检测。结果显示,Pornd1 c DNA开放阅读框为699 bp,编码232个氨基酸。结合前期GWAS分析数据,本研究对Pornd1基因扩增和测序,确定Pornd1基因内含子2上存在一个抗迟缓爱德华氏菌病相关单核苷酸多态性位点,该位点在易感家系和抗病家系中分别是C/T,抗病家系(freqT=0.92)高于易感家系(freqT=0.20),具有显著性差异(P<0.05)。Pornd1在心、肝和肾中的相对表达量较高;迟缓爱德华氏菌感染后肝、肾和脾中Pornd1的表达量在6 h降低后逐渐升高,在48 h达到最高。Pornd1在抗病家系肝脏中的表达量显著高于易感家系。在蛋... 相似文献