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염색체의 배수성 증가에 의한 닭의 신품종 개발 1 . 다배수성 배아의 생산
여정수(J . S . Yeo),정선부(S . B . Chung),오봉국(B . K . Ohh),정일정(I . C . Cheong) 한국축산학회 1989 한국축산학회지 Vol.31 No.5
Tri-ethylene melamine (0.25-0.3㎎/2㎏ body wt.) which inhibits partially gametogenesis to produce the polyploid embryos was injected 4 hours before ovulation to dual purpose chicken with 24 hours egg cycle. After artificial insemination 78-83% of fertilization rate and 44% of triploid embryos of fertilized eggs were observed. Times of meiosis I and Q during oogenesis detected by the sex chromosomes of triploid embryos seemed to be at 2-4 hours before ovulation and duration meiosis Ⅱ was shorter and more irregular than that of meiosis I.
Kasthuri, S.R.,Wan, Q.,Whang, I.,Lim, B.S.,Yeo, S.Y.,Choi, C.Y.,Lee, J. Academic Press 2014 Fish & Shellfish Immunology Vol.40 No.2
Antimicrobial immune defense is evolutionarily preserved in all organisms. Mammals have developed robust, protein-based antiviral defenses, which are under constant investigation. Studies have provided evidences for the various fish immune factors sharing similarity with those of mammals. In this study, we have identified an ortholog of mitochondrial antiviral signaling protein from rock bream, Oplegnathus fasciatus. RbMAVS cDNA possesses an open reading frame (ORF) of 1758 bp coding for a protein of 586 amino acids with molecular mass of approximately 62 kDa and isoelectric point of 4.6. In silico analysis of RbMAVS protein revealed a caspase recruitment domain (CARD), a proline rich domain and a transmembrane domain. RbMAVS protein also contains a putative TRAF2 binding motif, <SUP>319</SUP>PVQDT<SUP>323</SUP>. Primary sequence comparison of RbMAVS with other orthologues revealed heterogeneity towards the C-terminus after the CARD region. RbMAVS transcripts were evident in all the examined tissues. RbMAVS expression was induced in vivo after poly I:C challenge in peripheral blood cells, liver, head kidney and spleen tissues. Over-expression of RbMAVS potently inhibited marine birnavirus (MABV) infection in rock bream heart cells and induced various cytokines and signaling molecules in vitro. Thus, RbMAVS is an antiviral protein and potentially involved in the recognition and signaling of antiviral defense mechanism in rock bream.
Elvitigala, D.A.S.,Whang, I.,Premachandra, H.K.A.,Umasuthan, N.,Oh, M.J.,Jung, S.J.,Yeo, S.Y.,Lim, B.S.,Lee, J.H.,Park, H.C.,Lee, J. Academic Press 2012 Fish & Shellfish Immunology Vol.33 No.1
Caspase 3 is a prominent mediator of apoptosis and participates in the cell death signaling cascade. In this study, caspase 3 was identified (Rbcasp3) and characterized from rock bream (Oplegnathus fasciatus). The full-length cDNA of Rbcasp3 is 2683 bp and contains an open reading frame of 849 bp, which encodes a 283 amino acid protein with a calculated molecular mass of 31.2 kDa and isoelectric point of 6.31. The amino acid sequence resembles the conventional caspase 3 domain architecture, including crucial amino acid residues in the catalytic site and binding pocket. The genomic length of Rbcasp3 is 7529 bp, and encompasses six exons interrupted by five introns. Phylogenetic analysis affirmed that Rbcasp3 represents a complex group in fish that has been shaped by gene duplication and diversification. Many putative transcription factor binding sites were identified in the predicted promoter region of Rbcasp3, including immune factor- and cancer signal-inducible sites. Rbcasp3, excluding the pro-domain, was expressed in Escherichia coli. The recombinant protein showed a detectable activity against the mammalian caspase 3/7-specific substrate DEVD-pNA, indicating a functional role in physiology. Quantitative real time PCR assay detected Rbcasp3 expression in all examined tissues, but with high abundance in blood, liver and brain. Transcriptional profiling of rock bream liver tissue revealed that challenge with lipopolysaccharides (LPS) caused prolonged up-regulation of Rbcasp3 mRNA whereas, Edwardsiella tarda (E. tarda) stimulated a late-phase significant transcriptional response. Rock bream iridovirus (RBIV) up-regulated Rbcasp3 transcription significantly at late-phase, however polyinosinic-polycytidylic acid (poly(I:C)) induced Rbcasp3 significantly at early-phase. Our findings suggest that Rbcasp3 functions as a cysteine-aspartate-specific protease and contributes to immune responses against bacterial and viral infections.
Lee, Y.,De Zoysa, M.,Whang, I.,Lee, S.,Kim, Y.,Oh, C.,Choi, C.Y.,Yeo, S.Y.,Lee, J. Academic Press 2011 Fish & Shellfish Immunology Vol.30 No.2
The caspase family represents aspartate-specific cysteine proteases that play key roles in apoptosis and immune signaling. In this study, we cloned the first death effector domain (DED)-containing molluscan caspase-8 gene from disk abalone (Haliotis discus discus), which is named as hdCaspase-8. The full-length hdCaspase was 2855 bp, with a 1908 bp open reading frame encoding 636 amino acids. The hdCaspase-8 had 72 kDa predicted molecular mass with an estimated isoelectric point (PI) of 6.0. The hdCaspase-8 amino acid sequence contained the characteristic feature of an N-terminal two DED, a C-terminal catalytic domain and the caspase family cysteine active site <SUP>513</SUP>KPKLFFLQACQG<SUP>524</SUP>. Phylogenetic analysis results showed that hdCaspase-8 is more similar to the invertebrate Tubifex tubifex (sludge worm) caspase-8. Real-time RT-PCR results showed that hdCaspase-8 constitutively and ubiquitously expressed in all tested tissue of unchallenged disk abalone. The basal expression level of hdCaspase-8 in gill tissue was higher than all other tested tissues. The hdCaspase-8 mRNA expression in gill and hemocytes was significantly up-regulated by exposure to bacteria (Vibrio alginolyticus, Vibrio parahemolyticus and Listeria monocytogenes) and VHSV (viral hemorrhagic septicemia virus), as compared to control animals. These results suggest that hdCaspase-8 may be involved in immune response reactions in disk abalone.
Premachandra, H.K.A.,Elvitigala, D.A.S.,Whang, I.,Kim, E.,De Zoysa, M.,Lim, B.S.,Yeo, S.Y.,Kim, S.,Park, M.A.,Park, H.C.,Lee, J. Academic Press 2013 Fish & shellfish immunology Vol.34 No.6
Cystatins are a well-characterized group of cysteine protease inhibitors, which play crucial roles in physiology and immunity. In the present study, an invertebrate ortholog of cystatin B was identified in Manila clam (Ruditapes philippinarum) (RpCytB) and characterized at the molecular level, demonstrating its inhibitory activity against the well-known cysteine protease, papain. The complete coding sequence of RpCytB (297 bp in length) encodes a 99 amino acid peptide with a calculated molecular mass of 11 kDa and a theoretical isoelectric point of 5.9. The derived peptide was found to harbor typical features of cystatin proteins, including the 'Q-X-V-X-G' motif, which was identified as QLVAG in RpCytB. Phylogenetic analysis of RpCytB revealed close evolutionary relationships with its invertebrate counterparts, especially those from mollusks. Recombinant RpCytB (rRpCytB) was overexpressed as a fusion with maltose binding protein (MBP) in Escherichia coli BL21 (DE3) cells. Purified rRpCytB fusion protein exhibited a detectable inhibitory activity against papain, while the control MBP showed an almost constant negligible activity. While quantitative RT-PCR detected ubiquitous RpCytB expression in all tissues examined, the expressions in hemocytes and gills were relatively higher. Upon in vivo immune challenge with lipopolysaccharide (LPS), the expression of RpCytB in gills and hemocytes was down-regulated. Similar challenges with poly I:C and intact Vibrio tapetis bacteria revealed a complicated transcriptional regulation, wherein mRNA expression levels fluctuated over time of exposure. Moreover, a precise induction of RpCytB expression after bacterial infection was detected in gills by in situ hybridization. Collectively, our findings in this study indicate that RpCytB expression is sensitive to host pathological conditions and may contribute cysteine protease inhibitory activity to modulate the immune response.
Elvitigala, D.A.S.,Premachandra, H.K.A.,Whang, I.,Priyathilaka, T.T.,Kim, E.,Lim, B.S.,Jung, H.B.,Yeo, S.Y.,Park, H.C.,Lee, J. Academic Press 2013 Fish & shellfish immunology Vol.35 No.4
Catalases are well known antioxidant enzymes that can mainly dismutate hydrogen peroxide into water and oxygen in order to prevent oxidative stress. The complete genomic DNA (gDNA) sequence of the catalase gene from rock bream (Oplegnathus fasciatus) was identified from our custom-constructed BAC genomic DNA library and designated as RbCat. RbCat consists of 13 exons, separated by 12 introns, within a 13,722-bp gDNA sequence. The complete cDNA sequence (3303 bp) of RbCat is comprised of a 1581-bp coding region, encoding a peptide of 527 amino acids (aa) in length, with a predicted molecular mass of 60 kDa and a theoretical isoelectric point of 8.34. The anticipated promoter region of RbCat contains several transcription factor-binding sites, including sites that bind with immune- and antioxidant-responsive signaling molecules, suggesting its substantial transcriptional regulation. RbCat resembles the typical catalase family signature, i.e., it is composed of the catalase proximal active site motif along with a catalase proximal heme-ligand signature motif and shares great homology with its fish counterparts. According to multiple sequence alignment, functionally important amino acids present in RbCat were thoroughly conserved among its vertebrate counterparts. Phylogenetic analysis revealed that RbCat evolved from a vertebrate origin, and further positioned it in the fish clade. Recombinant RbCat had noticeable peroxidase activity against its substrate, hydrogen peroxide, in a dose-dependent manner. However, it demonstrated substantial peroxidase activity within a broad range of temperatures and pH values. Constitutive RbCat mRNA expression of different magnitudes was detected in a tissue-specific manner, suggesting its diverse role in physiology with respect to the tissue type. Moreover, immune challenge experiments using Edwardsiella tarda and rock bream iridovirus (RBIV) as live pathogens and polyinosinic:polycytidylic acid and lipopolysaccharide as mitogens revealed that the transcription of RbCat can be modulated by immune stimulation. Collectively, the results obtained in this study suggest that RbCat can function as a potent antioxidant enzyme in rock bream and may play a role in post-immune responses with respect to its peroxidase activity.
Yeo, I. C.,Lee, N. K.,Yang, B. W.,Hahm, Y. T. HUMANA PRESS INC 2014 Applied biochemistry and biotechnology Vol.172 No.2
Bacillus subtilis SC-8 produces an antibiotic that has narrow antagonistic activity against bacteria in the Bacillus cereus group. In B. cereus group bacteria, peptide-activating PlcR (PapR) plays a significant role in regulating the transcription of virulence factors. When B. subtilis SC-8 and B. cereus are co-cultured, PapR is assumed to stimulate antibiotic production by B. subtilis SC-8. To better understand the effect of PapR on this interspecies interaction, the global transcriptome profile of B. subtilis SC-8 was analyzed in the presence of PapR. Significant changes were detected in 12.8 % of the total transcripts. Genes related to amino acid transport and metabolism (16.5 %) and transcription (15 %) were mainly upregulated, whereas genes involved in carbohydrate transport and metabolism (12.7 %) were markedly downregulated. The expression of genes related to transcription, including several transcriptional regulators and proteins involved in tRNA biosynthesis, was increased. The expression levels of genes associated with several transport systems, such as antibiotic, cobalt, and iron complex transporters, was also significantly altered. Among the downregulated genes were transcripts associated with spore formation, the subtilosin A gene cluster, and nitrogen metabolism.
Measurement of the refractive index of the LAB-based liquid scintillator and acrylic at RENO
Yeo, I S,Jang, J H,Kim, M S,Ahn, J K,Baik, S R,Choi, E I,Choi, Seonho,Choi, Y,Jang, H I,Jang, J S,Jeon, E J,Jeong, I S,Joo, K K,Kim, B C,Kim, H S,Kim, J Y,Kim, S B,Kim, S H,Kim, W,Kim, Y D,Lee, J,Lim, Royal Swedish Academy of Sciences 2010 Physica scripta Vol.82 No.6
<P>In this paper, we describe the first measurement of the refractive index of the linear alkyl benzene (LAB)-based liquid scintillator and acrylic used at the Reactor Experiment for Neutrino Oscillation (RENO). The refractive index of a material is an important optical property and the prism spectrometer is the most common device for measuring the refractive index. Using the minimum deviation technique, which requires an exact measurement of the apex angle, we determined the refractive indices at six different wavelengths and then fitted them. We also compared the refractive index of the LAB-based liquid scintillator with the refractive indices of other liquid scintillator solvents commonly used in reactor neutrino experiments.</P>