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1.
Proteins of the DYRK(dual-specificity tyrosine-phosphorylation-regulated kinase) family are characterized by the presence of a conserved kinase domain and N-terminal DH box.DYRK2 is involved in regulating key developmental and cellular processes,such as neurogenesis,cell proliferation,cytokinesis,and cellular differentiation.Herein,we report that the ortholog of DYRK2 found in zebrafish shares about 70% identity with that of human,mouse,and chick.RT-PCR showed that DYRK2 is expressed maternally and zygotica... 相似文献
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We extracted marine low-temperature lysozyme (MLTL), a novel lysozyme, from a marine microorganism through fermentation. Our
previous study suggested that a low molecular weight (16 kDa) may exert anti-tumor activity through antiangiogenesis. In this
study, we extracted a high weight (39 kDa) and investigated its antiangiogenic activity in vivo and in vitro. Using zebrafish embryos as an in vivo study model, we found that treatment with MLTL significantly inhibited the growth of subintestinal vessels (SIVs) in a dose-dependent
manner and that 400 μg/ml MLTL was sufficient to block the growth of SIVs. An in vitro study conducted using human umbilical vein endothelial cells (HUVECs) revealed that MLTL suppressed the proliferation, migration
and tube formation of HUVECs in a dose-dependent manner. Interestingly, assays by flow cytometry and DNA electrophoresis indicated
that MLTL was able to induce apoptosis of HUVECs. Moreover, further study demonstrated that the disruption of intracellular
Ca2+ homeostasis may play an important role in MLTL induced apoptosis of HUVECs. Taken together, the results of this study demonstrate
for the first time that MLTL inhibits angiogenesis through its pleiotropic effects on vascular endothelial cells and induces
apoptosis through regulation of cellular Ca2+ levels. The results of this study also revealed a possible mechanism underlying the antiangiogenic effect of MLTL and suggested
that MLTL may be a promising new antiangiogenic agent for use in cancer therapy. 相似文献
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Shuo Li Chunmei Li Xubo Wang Yanan Wang Zhipeng Liu Teng Zhai Quanqi Zhang 《中国海洋大学学报(英文版)》2010,9(1):59-64
A novel immune-related gene was expressed in Japanese flounder (Paralichthys olivaceus) injected with Vibrio anguillarum. The complete cDNA contained a 169 bp 5’UTR, a 336 bp open reading frame (ORF) encoding 111 amino acids and a 556bp 3’UTR.
Six exons and five introns were identified in the PoIR2 gene. Blastp similarity comparison showed its encoding protein had 50% similarity to Danio rerio neuromedin S (NMS), but further alignment indicated they did not have NMS C-terminal conservational signature domain. So
it was not defined as an NMS homologue. Protein structure analysis indicated it had a 26aa signal peptide and was a secretory
pathway protein. RT-PCR demonstrated that the expression of PoIR2 was quickly induced and drastically increased in liver, kidney, spleen, gills, intestine, heart, and skeletal muscle after
infected with V. anguillarum. These results indicated that the PoIR2 might play some important role in Japanese flounder immune response system. This gene was named PoIR2 (P.olivaceus immune-related gene 2, GenBank accession number: EU224372). The mature PoIR2 peptide was expressed in BL21(DE3) pLysS using pET-32a(+) vector and a great part of the recombinant mature peptide existed
as soluble type. 相似文献
6.
To improve the expression efficiency of exogenous genes in Chlamydomonas reinhardtii, a high efficient expression vector was constructed. Green fluorescent protein （GFP） was expressed in C. reinhardtii under the control of promoters： RBCS2 and HSP70A-RBCS2. Efficiency of transformation and expression were compared between two transgenic algae： RBCS2 mediated strain Tran-Ⅰ and HSP70A-RBCS2 mediated strain Tran-Ⅱ. Results show that HSP70A-RBCS2 could improve greatly the transformation efficiency by approximately eightfold of RBCS2, and the expression efficiency of GFP in Tran-Ⅱ was at least double of that in Tran-Ⅰ. In addition, a threefold increase of GFP in Tran-Ⅱ was induced by heat shock at 40℃. All of the results demonstrated that HSP70A-RBCS2 was more efficient than RBCS2 in expressing exogenous gene in C. reinhardtii. 相似文献
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MyoD, Myf5, and myogenin are myogenic regulatory factors that play important roles during myogenesis. It is thought that MyoD and Myf5 are required for myogenic determination, while myogenin is important for terminal differentiation and lineage maintenance. To better understand the function of myogenic regulatory
factors in muscle development of flounder, an important economic fish in Asia, real-time quantitative RT-PCR was used to characterize
the expression patterns of MyoD, Myf5, and myogenin at early stages of embryo development, and in different tissues of the adult flounder. The results show that, Myf5 is the first gene to be expressed during the early stages of flounder development, followed by MyoD and myogenin. The expressions of Myf5, yoD, and myogenin at the early stages have a common characteristic: expression gradually increased to a peak level, and then gradually decreased
to an extremely low level. In the adult flounder, the expression of the three genes in muscle is much higher than that in
other tissues, indicating that they are important for muscle growth and maintenance of grown fish. During embryonic stages,
the expression level of MyoD might serve an important role in the balance between muscle cell differentiation and proliferation. When the MyoD expression is over 30% of its highest level, the muscle cells enter the differentiation stage. 相似文献
8.
Expression of putative zinc-finger protein lcn61 gene in lymphocystis disease virus China （LCDV-cn） genome 总被引：1，自引：0，他引：1
An open reading frame (lcn61) of lymphocystis disease virus China (LCDV-cn), probably responsible for encoding putative zinc-finger proteins was amplified
and inserted into pET24a (+) vector. Then it expressed in E. coli BL21 (DE3), and His-tag fusion protein of high yield was obtained. It was found that the fusion protein existed in E. coli mainly as inclusion bodies. The bioinformatics analysis indicates that LCN61 is C2H2 type zinc-finger protein containing
four C2H2 zinc-finger motifs. This work provides a theory for functional research of lcn61 gene.
Supported by High Technology Research and Development Program of China (863 Program, No. 2006AA100309) 相似文献
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We studied the effects of mercury (Hg2+) on antioxidant and digestive enzyme activities in terms of LC50 value and on hepatopancreas histostructures of juvenile Chinese mitten crabs Eriocheir sinensis in 40-day exposure to various concentrations of Hg2+ (0, 0.01, 0.05, 0.10, 0.20, and 0.30 mg/L). The results show that the activities of superoxide dismutase (SOD), glutathione
peroxidase (GPX) and catalase (CAT) significantly increased in the concentrations of 0.01 and 0.05 mg/L, while that of enzyme
decreased in 0.10, 0.20 and 0.30 mg/L treatments. Meanwhile, Hg2+ disrupted the histostructures of the hepatopancreas, causing decreases in activities of pepsin, tryptase, amylase, and cellulose,
which are synthesized in the hepatopancreas. Moreover, as the Hg2+ concentration increased, the survival rate of the crabs decreased, worst at 56.57% in 0.30 mg/L. Therefore, although crabs
are able to tolerate low levels of mercury pollution, high levels lead to cellular injury and tissue damage in hepatopancreas,
which then loses some of its vital physiological functions such as absorption, storage, and secretion. 相似文献
10.
In 2008, a green tide broke out before the sailing competition of the 29th Olympic Games in Qingdao. The causative species
was determined to be Enteromorpha prolifera (Ulva prolifera O. F. Müller), a familiar green macroalga along the coastline of China. Rapid accumulation of a large biomass of floating
U. prolifera prompted research on different aspects of this species. In this study, we constructed a nonnormalized cDNA library from the
thalli of U. prolifera and acquired 10 072 high-quality expressed sequence tags (ESTs). These ESTs were assembled into 3 519 nonredundant gene groups,
including 1 446 clusters and 2 073 singletons. After annotation with the nr database, a large number of genes were found to
be related with chloroplast and ribosomal protein, GO functional classification showed 1 418 ESTs participated in photosynthesis
and 1 359 ESTs were responsible for the generation of precursor metabolites and energy. In addition, rather comprehensive
carbon fixation pathways were found in U. prolifera using KEGG. Some stress-related and signal transduction-related genes were also found in this study. All the evidences displayed
that U. prolifera had substance and energy foundation for the intense photosynthesis and the rapid proliferation. Phylogenetic analysis of
cytochrome c oxidase subunit I revealed that this green-tide causative species is most closely affiliated to Pseudendoclonium akinetum (Ulvophyceae). 相似文献
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The molecular basis of color polymorphism in the shells of the pearl oyster Pinctada fucata is largely unknown. We developed a red-shelled family line and used suppression subtractive hybridization (SSH) to screen for differentially expressed genes in red- and non-red-shelled pearl oysters. We constructed forward and reverse cDNA subtractive libraries consisting of 2 506 and 797 clones, respectively. Among 343 randomly selected clones in the forward library, 304 sequences were identified in GenBank using BLASTx and BLASTn. Of the 304 sequences, 13 showed no similarity to known sequences and 291 were matched with known genes of the pearl oyster, including shematrin-1, shematrin-2, shematrin-6, shematrin-7, nacrein, nacrein-like protein, aspein for shell matrix protein, glycine-rich protein, mantle gene 5, 28S, EST00031, EST00036, 16S, and COΙ. In the reverse library, 7 clones were sequenced and analyzed by BLAST. Two sequences shared similarity with EST00036 from the P. fucata subtraction cDNA library, four with the P. fucata mitochondrial gene for 16S rRNA and 1 with P. fucata shematrin-2. We evaluated the expression of 12 genes from the forward library using RT PCR. Two sequences matched with 16S and COΙ so were considered to be false positives. The remaining 10 sequences were differentially expression in the red-shelled pearl oysters. Our results suggest that differential expression of these genes may be related to color variation in the red-shelled family line of the pearl oyster. 相似文献
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Estradiol, or 17-β-estradiol (E2), the most potent naturally occurring estrogen, is involved in the hormone-immune system interaction in both mammals and
fish. However, in vivo studies are largely limited, and little is known about whether E2 exerts similar effects on both female and male zebrafish (Danio rerio). Here, we show exposure of both sexes of D. rerio to 20 nmol/L E2 resulted in a significant increase in Vg1 expression, but caused little damage to the hepatocytes, suggesting that this is the optimum E2 concentration. Also, exposure to 20 nmol/L E2 for 20 days caused a marked increase in plasma IgM levels, but had little influence on the peripheral leukocyte density,
providing the first evidence of a hormone-immune system interaction in this species. 相似文献
13.
Zhigao Sun Xiaojie Mou Jingkuan Sun Hongli Song Xiang Yu Lingling Wang Huanhuan Jiang Wanlong Sun Wenguang Sun 《中国地理科学(英文版)》2012,22(1):15-28
From April 2008 to November 2009,the nitrogen(N) cycle of plantsoil system in seepweed(Suaeda salsa) wetland in the intertidal zone of the Huanghe(Yellow) River estuary was studied.Results showed that soil N had sig-nificant seasonal fluctuations and vertical distribution,and the net N mineralization rates in topsoil were significantly different in growing season(p < 0.01).The N/P ratio(9.87 ± 1.23) of S.salsa was less than 14,indicating that plant growth was limited by N.The N accumulated in S.salsa litter at all times during decomposition,which was ascribed to the N immobilization by microbes from the environment.Soil organic N was the main N stock of plant-soil system,accounting for 97.35% of the total N stock.The N absorption and utilization coefficients of S.salsa were very low(0.0145 and 0.3844,respectively),while the N cycle coefficient was high(0.7108).The results of the N turnovers among compartments of S.salsa wetland showed that the N uptake amount of aboveground part and root were 7.764 g/m2and 4.332 g/m2,respectively.The N translocation amounts from aboveground part to root and from root to soil were 3.881 g/m2 and 0.626 g/m2,respectively.The N translocation amount from aboveground living body to litter was 3.883 g/m2,the annual N return amount from litter to soil was more than 0.125(-) g/m2(minus represented immobilization),and the net N mineralization amount in topsoil(0-15 cm) in growing season was 1.190 g/m2.The assessment of N biological cycle status of S.salsa wetland indicated that N was a very important limiting factor and the ecosystem was situated in unstable and vulnerable status.The S.salsa was seemingly well adapted to the low-nutrient status and vulnerable habitat,and the N quantitative relationships determined in the compartment model might provide scientific base for us to reveal the special adaptive strategy of S.salsa to the vulnerable habitat in the following studies. 相似文献
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The tolerance to ammonia and nitrites in freshwater ciliate Paramecium bursaria was measured in a conventional open system. The ciliate was exposed to different concentrations of ammonia and nitrites for 2h and 12h in order to determine the lethal concentrations. Linear regression analysis revealed that the 2h-LC50 value for ammonia was 95.94 mg/L and for nitrite 27.35 mg/L using probit scale method (with 95% confidence intervals). There was a linear correlation between the mortality probit scale and logarithmic concentration of ammonia which fit by a regression equation y=7.32x-9.51 (R^2=0.98; y, mortality probit scale; x, logarithmic concentration of ammonia), by which 2 h-LC50 value for ammonia was found to be 95.50 mg/L. A linear correlation between mortality probit scales and logarithmic concentration of nitrite is also followed the regression equation y=2.86x 0.89 (R^2=0.95; y, mortality probit scale; x, logarithmic concentration of nitrite). The regression analysis of toxicity curves showed that the linear correlation between exposed time of ammonia-N LC50 valueand ammonia-N LC50 value followed the regression equation y=2 862.85e-0.08x (R^2=0.95; y, duration of exposure to LC50 value; x, LC50 value), and that between exposed time of nitrite-N LC50 value and nitrite-N LC50 value followed the regression equation y = 127.15e-0.13x (R^2=0.91; y, exposed time of LC50 value; x, LC50 value). The results demonstrate that the tolerance to ammonia in P. bursaria is considerably higher than that of the larvae or juveniles of some metozoa, e.g. cultured prawns and oysters. In addition, ciliates, as bacterial predators, are likely to play a positive role in maintaining and improving water quality in aquatic environments with high-level ammonium, such as sewage treatment systems. 相似文献
15.
Proteomics was used to reveal the differential protein expression profiles of acute responses to copper sulfate exposure in
larvae of Artemia sinica. Fourteen differentially displayed protein spots were detected and seven of them were identified. Three spots were up-expressed
and identified: actin, heat shock protein 70, and chaperone subunit 1; three down-regulated proteins were identified: arginine
kinase, elongation factor-2, and glycine-rich protein; and a newly expressed protein was identified as peroxiredoxin. The
study indicates the involvement of all the differentially expressed proteins in the early responses of protein expression,
and in the survival of A. sinica in the presence of copper and other heavy metals; the findings improve understanding of the organism’s adaptive responses
and resistance. 相似文献
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Insulin-like growth factors (IGFs) are key regulators of development and growth. Here, we characterized the igf2 gene from olive flounder (Paralichthys olivaceus) and determined its temporal and spatial expression. We set up an in-vitro protein expression system in eukaryotic human embryonic kidney (HEK293T) cells and explored its effects on cell proliferation. The flounder igf2 cDNA contained a 648-bp open reading frame (ORF) encoding a protein of 215 amino acids (aa), which spanned the complete signal peptide (47 aa), mature peptide (70 aa), and E domain (98 aa). In adult flounder, igf2 mRNA was detected in all selected tissues. In early development, igf2 mRNA was detected throughout development from unfertilized eggs to hatching-stage embryos. In-situ hybridization analysis indicated that igf2 mRNA was specially expressed in the brain region, floor plate, hypochord, otic vesicle, and pectoral fin during embryogenesis. Western blotting analysis indicated that the soluble recombinant flounder IGF2 protein was successfully produced through eukaryotic expression in HEK293T cells. In addition, the recombinant IGF2 protein significantly promoted the proliferation of human cervical carcinoma (HeLa) and HEK293T cells. These results provide new information about the structural and functional conservation, expression patterns, and biological activity of the igf2 in teleosts.
相似文献17.
Lymphocystis disease, caused by the lymphocystis disease virus (LCDV), is a significant worldwide problem in fish industry
causing substantial economic losses. In this study, we aimed to develop the DNA vaccine against LCDV, using DNA vaccination
technology. We evaluated plasmid pEGFP-N2-LCDV1.3 kb as a DNA vaccine candidate. The plasmid DNA was transiently expressed
after liposome transfection into the eukaryotic COS 7 cell line. The distribution and expression of the DNA vaccine (pEGFP-N2-LCDV1.3kb)
were also analyzed in tissues of the vaccinated Japanese flounder by PCR, RT-PCR and fluorescent microscopy. Results from
PCR analysis indicated that the vaccine-containing plasmids were distributed in injected muscle, the muscle opposite the injection
site, the hind intestine, gill, spleen, head, kidney and liver, 6 and 25 days after vaccination. The vaccine plasmids disappeared
100 d post-vaccination. Fluorescent microscopy revealed green fluorescence in the injected muscle, the muscle opposite the
injection site, the hind intestine, gill, spleen, head, kidney and liver of fish 48 h post-vaccination, green fluorescence
did not appear in the control treated tissue. Green fluorescence became weak at 60 days post-vaccination. RT-PCR analysis
indicated that the mcp gene was expressed in all tested tissues of vaccinated fish 6–50 days post-vaccination. These results demonstrate that the
antigen encoded by the DNA vaccine is distributed and expressed in all of the tissues analyzed in the vaccinated fish. The
antigen would therefore potentially initiate a specific immune response. the plasmid DNA was injected into Japanese flounder
(Paralichthys olivaceus) intramuscularly and antibodies against LCDV were evaluated. The results indicate that the plasmid encoded DNA vaccine could
induce an immune response to LCDV and would therefore offer immune protection against LCD. Further studies are required for
the development and application of this promising DNA vaccine. 相似文献
18.
Information regarding immunocompetence of the adaptive immune system(AIS) in zebrafish Danio rerio remains limited.Here,we stimulated an immune response in fish embryos,larvae and adults using lipopolysaccharide(LPS) and measured the upregulation of a number of AIS-related genes(Rag2,AID,TCRAC,IgLC-1,mIg,sIg,IgZ and DAB) 3 and 18 h later.We found that all of the genes evaluated were strongly induced following LPS stimulation,with most of them responding at 8 d post fertilization.This confirms that a functional adaptive immune response is present in D.rerio larvae,and provides a window for further functional analyses. 相似文献
19.
Ubiquitin, a highly conserved stress-related protein, is assigned multiple functions, such as DNA processing, protein degradation, and ribosome synthesis. The Crassostrea hongkongensis ubiquitin gene (designated ChUbL40) was cloned by a combination of suppressive subtractive hybridization (SSH) and rapid amplification of cDNA ends (RACE). The full-length cDNA of ChUbL40 is 496 bp in length, consisting of a 5’ untranslated region (UTR) of 34 bp, a 3’-UTR of 75 bp and an open reading frame of 387 bp encoding a ubiquitin fusion protein of 128 amino acids. Analysis of the amino acid sequence of ChUbL40 reveals that UbL40 is highly conservative during evolution. The expression patterns of ChUbL40 gene in various tissues were examined by real-time PCR. The expression level of ChUbL40 in haemocytes is down-regulated at 4 h and gradually returned to its original level from 6 h to 24 h after Vibrio alginolyticus challenge. Our results suggest that ChUbL40 is ubiquitously expressed and plays an important role in immune defense against bacterial challenge. 相似文献
20.
Nano-sized zinc oxide (nZnO) particles are one kind of the most commonly used metal oxide nanoparticles (NPs). This study compared the cytotoxic and embryotoxic effects of three increasing sized ZnO particles (? 30 nm, 80-150 nm and 2 μm) in the flounder gill (FG) cells and zebrafish embryos, and analyzed the contribution of size, agglomeration and released Zn2+ to the toxic effects. All the tested ZnO particles were found to be highly toxic to both FG cells and zebrafish embryos. They induced growth inhibition, LDH release, morphological changes and apoptosis in FG cells in a concentration-, size- and time-dependent manner. Moreover, the release of LDH from the exposed FG cells into the medium occurred before the observable morphological changes happened. The ultrasonication treatment and addition of serum favored the dispersion of ZnO particles and alleviated the agglomeration, thus significantly increased the corresponding cytotoxicity. The released Zn2+ ions from ZnO particles into the extracellular medium only partially contributed to the cytotoxicity. All the three sizes of ZnO particles tested induced developmental malformations, decrease of hatching rates and lethality in zebrafish embryos, but size- and concentration- dependent toxic effects were not so obvious as in FG cells possibly due to the easy aggregation of ZnO particles in freshwater. In conclusion, both FG cells and zebrafish embryos are sensitive bioassay systems for safety assessment of ZnO particles and the environmental release of ZnO particles should be closely monitored as far as the safety of aquatic organisms is concerned. 相似文献