• Title/Summary/Keyword: fish frame

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Expression Profiling of WSSV ORF 199 and Shrimp Ubiquitin Conjugating Enzyme in WSSV Infected Penaeus monodon

  • Jeena, K.;Prasad, K. Pani;Pathan, Mujahid Khan;Babu, P. Gireesh
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.8
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    • pp.1184-1189
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    • 2012
  • White spot syndrome virus (WSSV) is one of the major viral pathogens affecting shrimp aquaculture. Four proteins, WSSV199, WSSV 222, WSSV 249 and WSSV 403, from WSSV are predicted to encode a RING-H2 domain, which in presence of ubiquitin conjugating enzyme (E2) in shrimp can function as viral E3 ligase and modulate the host ubiquitin proteasome pathway. Modulation of host ubiquitin proteasome pathway by viral proteins is implicated in viral pathogenesis. In the present study, a time course expression profile analysis of WSSV Open Reading Frame (ORF) 199 and Penaeus monodon ubiquitin conjugating enzyme (PmUbc) was carried out at 0, 3, 6, 12, 24, 48 and 72 h post WSSV challenge by semi-quantitative RT-PCR as well as Real Time PCR. EF1${\alpha}$ was used as reference control to normalize the expression levels. A significant increase in PmUbc expression at 24 h post infection (h.p.i) was observed followed by a decline till 72 h.p.i. Expression of WSSV199 was observed at 24 h.p.i in WSSV infected P. monodon. Since the up-regulation of PmUbc was observed at 24 h.p.i where WSSV199 expression was detected, it can be speculated that these proteins might interact with host ubiquitination pathway for viral pathogenesis. However, further studies need to be carried out to unfold the molecular mechanism of interaction between host and virus to devise efficient control strategies for this chaos in the shrimp culture industry.

Molecular Cloning, Characterization and Expression Analysis of an ILF2 Homologue from Tetraodon nigroviridis

  • Wang, Hui-Ju;Shao, Jian-Zhong;Xiang, Li-Xin;Shen, Jia
    • BMB Reports
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    • v.39 no.6
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    • pp.686-695
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    • 2006
  • Interleukin-2 enhancer binding factor 2 (ILF2) was reported to regulate transcription of interleukin-2 (IL-2), a central cytokine in the regulation of T-cell responses. This property of ILF2 was well characterized in human and mammals, but little is known in bony fish. In this paper, an ILF2 homologue was cloned and well characterized from Tetraodon nigrovirid is for the further investigation of the function of ILF2 in bony fish. The full-length Tetraodon ILF2 cDNA was 1380 bp in size and contained an open reading frame (ORF) of 1164 bp that translates into a 387 amino-acid peptide with a molecular weight of 42.9 kDa, a 5' untranslated region (UTR) of 57 bp, and a 3' UTR of 159 bp containing a poly A tail. The deduced peptide of Tetraodon ILF2 shared an overall identity of 58%~93% with other known ILF2 sequences, and contained two N-glycosylation sites, two N-myristoylation sites, one RGD cell attachment sequence, six protein kinase C phosphorylation sites, one amino-terminal RGG-rich single-stranded RNA-binding domain, and a DZF zinc-finger nucleic acid binding domain, most of which were highly conserved through species compared. Constitutive expression of Tetraodon ILF2 was observed in all tissues examined, including gill, gut, head kidney, spleen, liver, brain and heart. The highest expression was detected in heart, followed by liver, head kidney and brain. Stimulation with LPS did not significantly alter the expression of Tetraodon ILF2. Gene organization analysis showed that the Tetraodon ILF2 gene have fifteen exons, one more than other known ILF2 genes in human and mouse. Genes up- and down-stream from the Tetraodon ILF2 were Rpa12, Peroxin-11b, Smad4, Snapap and Txnip homologue, which were different from that in human and mouse.

Molecular Cloning of Insulin-like Growth Factor-I (IGF-I) and IGF-II Genes of Marine Medaka (Oryzias dancena) and Their Expression in Response to Abrupt Transfer from Freshwater to Seawater

  • Kang, Yue-Jai;Kim, Ki-Hong
    • Fisheries and Aquatic Sciences
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    • v.13 no.3
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    • pp.224-230
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    • 2010
  • Growth hormone (GH) is known as one of the main osmoregulators in euryhaline teleosts during seawater (SW) adaptation. Many of the physiological actions of GH are mediated through insulin-like growth factor-I (IGF-I), and the GH/IGF-I axis is associated with osmoregulation of fish during SW acclimation. However, little information is available on the response of fish IGF-II to hyperosmotic stress. Here we present the first cloned IGF-I and IGF-II cDNAs of marine medaka, Oryzias dancena, and an analysis of the molecular characteristics of the genes. The marine medaka IGF-I cDNA is 1,340 bp long with a 257-bp 5' untranslated region (UTR), a 528 bp 3' UTR, and a 555-bp open reading frame (ORF) encoding a propeptide of 184 amino acid (aa) residues. The full-length marine medaka IGF-II cDNA consists of a 639 bp ORF encoding 212 aa, a 109 bp 5' UTR, and a 416 bp 3' UTR. Homology comparison of the deduced aa sequences with other IGF-Is and IGF-IIs showed that these genes in marine medaka shared high structural homology with orthologs from other teleost as well as mammalian species, suggesting high conservation of IGFs throughout vertebrates. The IGF-I mRNA level increased following transfer of marine medaka from freshwater (FW) to SW, and the expression level was higher than that of the control group, which was maintained in FW. This significantly elevated IGF-I level was maintained throughout the experiment (14 days), suggesting that in marine medaka, IGF-I is deeply involved in the adaptation to abrupt salinity change. In contrast to IGF-I, the increased level of marine medaka IGF-II mRNA was only maintained for a short period, and quickly returned a level similar to that of the control group, suggesting that marine medaka IGF-II might be a gene that responds to acute stress or one that produces a supplemental protein to assist with the osmoregulatory function of IGF-I during an early phase of salinity change.

Molecular Cloning and Characterization of the Estrogen Receptor from the Slender Bitterling (Acheilognathus yamatsutae)

  • Kim, Jong-Geuk;Kim, Ha-Ryong;Park, Yong-Joo;Chung, Kyu-Hyuck;Oh, Seung-Min
    • Environmental Analysis Health and Toxicology
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    • v.26
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    • pp.5.1-5.11
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    • 2011
  • Objectives: In order to identify the possibility of slender bitterling (SB) (Acheilognathus yamatsutae) being used as a test species for estrogenic endocrine disrupting chemicals (EEDCs), we carried out the cloning and sequence characterization of the estrogen receptor (ER). Methods: The ER from a slender bitterling was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR), 5'- and 3'-rapid amplification of cDNA ends (5'-RACE and 3'-RACE) and T-vector cloning. The expression of ER mRNA was also analyzed in six tissues (brain, liver, kidney, gill, gonad, and intestines) by real-time PCR. Results: We obtained an ER from the slender bitterling. The SB ER cDNA was 2189 base pairs (bp) in length and contained a 1707 bp open reading frame that encoded 568 amino acid residues. The SB ER amino acid sequence clustered in a monophyletic group with the $ER{\alpha}$ of other fish, and was more closely related to zebrafish $ER{\alpha}$(88% identity) than to the $ER{\alpha}$ of other fish. The SB ER cDNA was divided into A/B, C, D, E and F domains. The SB ER has conserved important sequences for ER functions, such as the DNA binding domain (D domain), which are consistent with those of other teleosts. Conclusions: The ER of the slender bitterling could provide basic information in toxicological studies of EEDCs in the slender bitterling.

Molecular identification and expression analysis of bactericidal permeability-increasing protein/ LPS-binding protein (BPI/LBP) from Black rockfish Sebastes schlegeli

  • Kwon, Mun-Gyeong;Kim, Ju-Won;Park, Myoung-Ae;Hwang, Jee-Youn;Park, Hyung-Jun;Baeck, Gun-Wook;Park, Chan-Il
    • Journal of fish pathology
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    • v.23 no.3
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    • pp.323-334
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    • 2010
  • Bactericidal/permeability-increasing protein (BPI) and lipopolysaccharide-binding protein (LBP) are important components of the mammalian innate defence system against Gram-negative infections. The BPI/LBP cDNA was identified from the black rockfish ConA/PMA or LPS stimulated leukocyte cDNA library. The full-length BR-BPI/LBP cDNA was 2118 bp long and contained an open reading frame (ORF) of 1422 bp that encoded 473 amino-acid residues. The 5' UTR had a length of 57 bp, and the 3' UTR 639 bp. The molecular weight and theoretical isoelectric point (pI) values were calculated 51.4 kDa and 9.72, respectively. Compared with other known BPI or BPI/LBP peptide sequences, the most conserved regions of the black rockfish BPI/LBP peptide were found to be the BPI1 N-terminal, BPI2 C-terminal domains and a LPS binding domain. Phylogenetic analysis based on the deduced amino acid sequence revealed a homologous relationship between the BPI/LBP sequence of black rockfish and that of other teleosts. The black rockfish BPI/LBP gene was predominantly expressed in the PBLs, head kidney, trunk kidney and spleen. The expression of the black rockfish BPI/LBP molecule was induced in the peripheral blood leukocytes (PBLs) from 1 to 24 h following LPS stimulation, with a peak at 12 h post-stimulation.

Handling Charactristics of FMT Net for the Larval Fish and Plankton (소형 어류 플랑크톤 채집용 FMT Net 의 운용 특성)

  • Jeong, Sun-Beom
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.41 no.2
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    • pp.122-128
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    • 2005
  • The experiments were conducted to fine out the handling characteristics of the FMT(Frame Mideater Trawl) in the southern waters of Korea using a trawler "DONGBAEK" belongs to Yosu National University. The realtionship between the net depth D(m) and the warp length L(m) at the towing speeds of 2.5k't and 3.5k't werw as follows ; D(m) = 0.30L - 1.3(2.5k't), D(m) = 0.16L - 1.5(3.5k't). Therefore, the net depth was 3.0m deeper when the warp length was 10m longer at the towing speed of 2.5k't and was 1.6m deeper for 10m longer at the speed of 3.5k't, respectively. The sinking speed of FMT was 6.5m/min when the warp releasing speed was 24m/min at the towing speed of 2.5k't and was 3.8m/min for 25m/min at the towing speed of 3.5k't, respectively. The rising speed of FMT was 6.9m/min when the warp rewinding speed was 28m/min at the towing speed of 2.5k't and was 5.3m/min for 25m/min at the towing speed of 3.5k't, respectively. The mean elapsed time getting to the stable towing condition was 104sec at the towing speed of 2.5k't and was 105sec at the towing speed of 3.5k't, respectively, and there was no time difference for the towing speed variation. During the towing, the net depth was comparatively stable on the condition of no change for the warp length and the towing speed.

Comparison on the Food Quality Characteristics of Muscles from Salmonids according to Species, Imported Country, and Separated Part (연어류 근육의 종류, 수입국 및 부위별 식품학적 품질 특성 비교)

  • Heu, Min Soo;Choi, Byeong Dae;Kim, Ki Hyun;Kang, Sang In;Kim, Yong Jung;Kim, Jin-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.1
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    • pp.16-25
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    • 2015
  • This study compared the food quality of salmonid fishes according to the species, country of origin, and separated part, such as fillet and frame. The proximate composition of chum salmon from Norway (CS-N) was 74.4% moisture, 19.5% crude protein, 4.2% crude lipid, and 1.2% ash. These values were within roughly 1% for the other salmon species. There was no significant difference (at P<0.05) in the Hunter a value of salmon muscle according to sepatated parts. However, there was a significant difference (P<0.05) in Hunter a value of salmon muscle according to the species and country of origin. There were significant differences in odor intensity and hardness of the salmon according to the species. The major free amino acid in all of the salmon muscles was anserine, which ranged from 61.3 to 73.0%. The taste value was the highest for salmon imported from Alaska (CS-A), followed by pink salmon, CS-N, and muscle separated from the frame (AS-C). In the taste value of all salmon muscles, the major amino acid was glutamic acid. The total amino acid content of salmon muscles ranged from 18.36 to 19.64 g/100 g, and the major amino acids were glutamic acid and aspartic acid. There were differences in the mineral contents, including Ca, P, K, and Fe, and fatty acid composition of salmon muscle according to species.

Structural Conservation and Food Habit-related Liver Expression of Uncoupling Protein 2 Gene in Five Major Chinese Carps

  • Liao, Wan-Qin;Liang, Xu-Fang;Wang, Lin;Fang, Ling;Lin, Xiaotao;Bai, Junjie;Jian, Qing
    • BMB Reports
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    • v.39 no.4
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    • pp.346-354
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    • 2006
  • The full-length cDNA of grass carp (Ctenopharyngodon idellus) and silver carp (Hypophthalmichthys molitrix) uncoupling protein 2 (UCP2) was obtained from liver. The grass carp UCP2 cDNA was determined to be 1152 bp in length with an open reading frame that encodes 310 amino acids. Five introns (Intron 3, 4, 5, 6 and 7) in the translated region, and partial sequence of Intron 2 in the untranslated region of grass carp UCP2 gene were also obtained. Gene structure comparison between grass carp and mammalian (human and mouse) UCP2 gene shows that, the UCP2 gene structure of grass carp is much similar to that of human and mouse. Partial UCP2 cDNA sequences of bighead carp (Aristichthys nobilis) and mud carp (Cirrhinus molitorella), were further determined. Together with the common carp (Cyprinus carpio) UCP2 sequence from GenBank (AJ243486), multiple alignment result shows that the nucleotide and amino acid sequences of the UCP2 gene, were highly conserved among the five major Chinese carps that belong to four subfamilies. Using beta-actin as control, the ratio UCP2/beta-actin mRNA (%) was determined to be $149.4{\pm}15.6$ (common carp), $127.4{\pm}22.1$ (mud carp), $96.7{\pm}12.7$ (silver carp), $94.1{\pm}26.8$ (bighead carp) and $63.7{\pm}16.2$ (grass carp). The relative liver UCP2 expression of the five major Chinese carps, shows a close relationship with their food habit: benthos and detrituseating fish (common carp and mud carp) > planktivorious fish (silver carp and bighead carp) > herbivorious fish (grass carp). We suggest that liver UCP2 might be important for Chinese carps to detoxify cyanotoxins and bacteria in debris and plankton food.

Design and performance evaluation of fish-luring system using the air-cooled LED lamp for jigging and angling boat (채낚기 어선용 공랭식 LED 집어시스템의 설계 및 성능평가)

  • Bae, Bong-Seong;Park, Byoung-Jae;Jeong, Eui-Cheol;Yang, Yong-Su;Park, Hae-Hoon;Chun, Young-Yull;Chang, Dae-Soo
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.45 no.2
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    • pp.85-95
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    • 2009
  • The fishing lamp is a fishing gear that gathers fish at night. But the cost of oil, which is used to light fishing lamp, has been risen significantly up to 30-40% of total fishing costs. Therefore it is very urgent to develop an energy economical fishing lamp in order to solve the business difficulties of fisheries. Under this background, this research aims at developing a fishing lamp for squid jigging and hairtail angling fishery using the LED, which has excellent energy efficiency and durability. The LED fishing lamp developed can be controlled to fix a fit direction of fish shoal deep because a fishing lamp can be adjustable up and down directions. One unit of fishing lamp has about an 80Watt capacity and the frame of fishing lamp is made of aluminium to emit generated heat of LED to outside. The LED lamp developed was highly durable, only 5.7% of emitting efficiency decreased for 18 months. The illuminance of a unit LED lamp was 2,070lux at 1m and 21lux at 10 m distance, and the intensity of LED lamp system emitted 2,580lux and 400lux at the respective distances. After development of this fishing lamp, 100 units are installed on operating fishing vessels. Experimental results show that energy consumption of squid jigging and hairtail angling was reduced by 40% and 87%, respectively. In conclusion, our methods showed elevated fishing power, compared with traditional fishing method: 37.7% for squid jigging and 24.5% for hairtail angling.

The Functional Relevance of Prepro-melanin Concentrating Hormone (pMCH) to Skin Color Change, Blind-side Malpigmentation and Feeding of Oliver Flounder Paralichthys olivaceus

  • Kang, Duk-Young;Kim, Hyo-Chan;Kang, Han-Seung
    • Fisheries and Aquatic Sciences
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    • v.17 no.3
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    • pp.325-337
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    • 2014
  • To assess the functional structure of prepro-melanin-concentrating hormone (pMCH), we isolated and cloned pMCH (of-pMCH) mRNA from the brain of the olive flounder, Paralichthys olivaceus, and compared its amino acid sequence with those from other animals. In addition, to examine whether activation of the brain of-pMCH gene is influenced by background color, density, and feeding, we compared pMCH mRNA activities against different background colors (bright and dark) and at different densities (100% PCA and 200% PCA). To examine whether the pMCH gene is related with malpigmentation of blind-side skin and appetite, we compared pMCH gene expression between ordinary and hypermelanic flounders, and between feeding and fasting flounders. The of-pMCH cDNA was 405 bp in the open reading frame [ORF] and encoded a protein of 135 amino acids; MCH was 51 bp in length and encoded a protein of 17 amino acids. An obvious single band of the expected size was obtained from the brain and pituitary by RT-PCR. In addition, of-pMCH gene activity was significantly higher in the bright background only at low density (< 100% PCA) making the ocular skin of fish whitening, and in ordinary fish. However, the gene activity was significantly decreased in dark background, at high density (>200% PCA), and in hypermelano fish. These results suggest that skin whitening camouflage of the flounder is induced by high MCH gene activity, and the density disturbs the function of background color in the physiological color change. Moreover, our data suggest that a low level of MCH gene activity may be related to malpigmentation of the blind-side skin. In feeding, although pMCH gene activity was significantly increased by feeding in the white background, the pMCH gene activity in the dark background was not influenced by feeding, indicating that the MCH gene activity increased by feeding can be offset by dark background color, or is unaffected by appetite. In conclusion, this study showed that MCH gene expression is related to ocular-skin whitening camouflage and blind-skin hypermelanosis, and is influenced by background color and density.