• Title/Summary/Keyword: Hybrid protein

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Suitable Dietary Protein/Lipid Ratio for Hybrid, Female Red Sea Bream Pagrus major and Male Black Sea Bream Acanthopagrus schlegeli in the Juvenile Stage, Compared with Red Sea Bream

  • Kim, Yang-Su;Ji, Seung-Cheol;Biswas, Amal;Biswas, Biswajit Kumar;Yong, Annita Seok Kian;Takaoka, Osamu;Jeong, Gwan-Sik;Murata, Osamu;Takii, Kenji
    • Fisheries and Aquatic Sciences
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    • v.17 no.1
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    • pp.75-84
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    • 2014
  • To determine a suitable dietary protein/lipid (CP/CL) ratio in the early juvenile stages of hybrid porgy ($F_1$), female red sea bream (RSB) ${\times}$ male black sea bream, five diets with various CP/CL ratios-60/7, 55/12, 51/17, 46/23, and 41/28-were prepared and provided to juveniles in triplicate. At the smaller juvenile stage, $F_1$, weighing 0.32 g, a significantly higher specific growth rate (SGR) and feed efficiency (FE) were seen with 60/7 and 55/12 diets. However, in RSB weighing 0.26 g, SGR and FE were higher with the 60/7 diet than the other diets at $21^{\circ}C$. At the larger juvenile stage, $F_1$, weighing 3.7 g, there was no significant difference in SGR or FE among the diets, but RSB weighing 4.0 g fed 60/7, 55/12, and 51/17 diets had higher SGR and FE than 46/23 and 41/28 diets at $24^{\circ}C$. Moreover, survival and apparent nutrient retention of $F_1$ at both stages were significantly higher than those in RSB. These results indicate that both $F_1$ and RSB weighing ca. 0.3 g require a higher dietary CP/CL than those weighing ca. 4 g. Additionally, $F_1$ in both trials showed the suitability of a lower dietary CP/CL than RSB, indicating that mass production of $F_1$ juveniles will be more economical than RSB.

A Study on the Effect of Physical Stimuli on Bone Cell Differentiation Using a Hybrid Bioreactor (Hybrid Bioreactor를 이용한 물리적 자극에 대한 세포반응 연구)

  • 이창양;최귀원;홍대희
    • Journal of Biomedical Engineering Research
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    • v.25 no.4
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    • pp.309-314
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    • 2004
  • in this study, hybrid bioreactor was used to apply physical stimuli in cell culture. Effect of the applied physical stimuli on the growth and differentiation of MC3T3-El cell in a three-dimensional Chitosan scaffold were studied by using the hybrid bioreactor. The hybrid bioreactor for physical stimulus was specially designed to apply uniaxial cyclic compressive and shear strain. Physical stimulus was applied over a period of 14 days with 150 cycles per day at a frequency of 0.5Hz. Strain magnitude was 2.5% of the scaffold size. Control group and physically stimulated group of the MC3T3-El tell were incubated and harvested at the indicated times (Day 6, 8, 10, 12, 14). The total amount of protein, which obtained information of cell growth, was determined by Lowey method. Alkaline phosphatase activity was examined by ELISA. Physically stimulated group using the hybrid bioreactor was increased in alkaline phosphatase activity comparing with control group. The nodule formation and calcium deposit of the physical stimuli group which resulted in cell differentiation was faster than that of control group.

Expression of Green Fluorescent Protein in Both Spodoptera frugiperda Cells and Bombyx mori Larvae by Ac-Bm Hybrid Virus

  • Jin, Byung-Rae;Yoon, Hyung-Joo;Yun, Eun-Young;Kang, Seok-Woo;Cho, Eun-Sook;Kang, Seok-Kwon
    • The Journal of Korean Society of Virology
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    • v.28 no.3
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    • pp.225-232
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    • 1998
  • We have expressed GFP in Sf9 and Bm5 cells or Bombyx mori larvae by using Ac-Bm hybrid virus capable of replicating in both Bm5 and Sf9 cells. Genomic DNA of Ac-Bm hybrid virus expressing ${\beta}$-galactosidase was cotransfected with baculovirus transfer vector containing GFP gene, pBacPAK-GFP in Sf9 cells. The Ac-Bm hybrid virus harboring GFP was named as Ac-Bm hybrid virus-GFP. The Ac-Bm hybrid virus-GFP-infected insect cells were easily selected by detecting the emission of GFP from each well of cell culture dish on the UV illuminator. GFP produced by Ac-Bm hybrid virus-GFP in Sf9 and Bm5 cells or B. mori larvae was confirmed by SDS-PAGE and Western blot analysis using GFP antibody. In addition, B. mori larvae infected with Ac-Bm hybrid virus-GFP was apparently appeared fluorescence from the whole body at S days postinoculation. The fluorescence of GFP from the hemolymph and fat body of B. mori larvae infected with Ac-Bm hybrid virus-GFP was also observed by fluorescence microscope. In conclusion, our results demonstrated that in baculovirus expression vector system, use of Ac-Bm hybrid virus have an additional advantage of expanded host range for producing recombinant proteins.

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Patterns of Protein Synthesis During the Second Cleavage of Mouse Two-Cell Embryos: Effects of Colcemid and a-Amanitin (생쥐 배아의 2세포기 분열과정에 있어서의 단백질 합성 분석 : Colcemid와 a-Amanitin의 영향)

  • Kang, Hae-Mook;Kvu
    • The Korean Journal of Zoology
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    • v.32 no.4
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    • pp.404-411
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    • 1989
  • In this study, we attempted to determine the precise patterns of protein synthesis during the second cleavage in mouse. F1 hybrid 2-cell embryos showing a highly synchronized cell cycle and outbreed ICR strain 2-cell embryos were used. The patterns of protein synthesis during the second cleavage showed the sequential changes in the F1 hybrid and ICR strain 2-cell embryos. Moreover, we examined the effects of mitotic and transcriptional inhibitors such as colcemid and a-amanitin on the protein synthesis in the late 2-cell embryos of ICR strain. Treatment of colcemid (0.1mg/ml) blocked the second cleavage, but did not affect on the change of protein synthesis. However, treatment of a-amanitin induced the synthesis of two set of polypeptides without affecting on synthesis of other proteins and cleavage. It thus seems that the appearance of a-amanitin-sensitive proteins may be not involved in the second cleavage. Therefore, these results indicate that the second cell cycle in mouse embryos appears to be regulated at post transcriptional level, presumably independent on the expression of embryonic genome. 본 연구는 생쥐 배아의 2세포기 분열과정중 단백질 합성양상과 단백질합성에 미치는 colcemid와 $\alpha$-amanitin의 영향을 조사하였다 이를 위하여 체내 수정된 ICR strain의 2세포기 배아와 매우 일치된 초기배아 분열양상을 보여주는 체외 수정된 F1 (C57BL x CBA) hybrid 2세포기 배아를 사용하였다. 두 종류의 2세포기 배아에서 단백질 합성은 분열단계에 따라서 매우 일치된 변화를 보여 주었다. 또한 유사분열 억제제인 colcemid (0.1mg/ml)의 처리는 2세포기 배아분열을 억제하였으나, 단백질 합성에는 아무런 변화를 주지 못하였다. 그리고 후기 2세포기 배아에 전사 억제제인 a-amanitin (100mg/ml)을 처리하였을 때 세포분열이나 다른 단백질의 합성에는 아무런 영향이 없이 단지 두개의 단백질의 합성만을 유도하였다. 이는 아마도 a-amanitin의 stress효과에 기인하는 것으로 추측된다. 따라서 생쥐 2세포기 배아의 분열과정은 배아게놈의 유전자 발현과는 무관하게 이미 합성되어 존재하는 mRNA에 의하여 조절되는 것으로 사료된다.

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Algorithm for extracting signaling pathways based on Protein-Protein Interaction and Protein location Information (Protein-Protein Interaction 에 세포 내 위치 정보를 활용한 단백질 신호전달 경로 추출 알고리즘 연구)

  • Jo, Mi-Kyung;Kim, Min-Kyung;Park, Hyun-Seok
    • Proceedings of the Korean Society of Computer Information Conference
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    • 2009.01a
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    • pp.77-84
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    • 2009
  • Intracellular signal transduction is achieved by protein-protein interaction. In this paper, we suggest performance algorithm based on Yeast protein-protein interaction and protein location information. We compare if pathways predicted with high valued weights indicate similar tendency with pathways provided in KEGG.

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Protein Interaction Databases and Its Application (단백질 상호작용 데이터베이스 현황 및 활용 방안)

  • Kim, Min Kyung;Park, Hyun Seok
    • IMMUNE NETWORK
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    • v.2 no.3
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    • pp.125-132
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    • 2002
  • In the past, bioinformatics was often regarded as a difficult and rather remote field, practiced only by computer scientists and not a practical tool available to biologists. However, the various on-going genome projects have had a serious impact on biological sciences in various ways and now there is little doubt that bioinformatics is an essential part of the research environment, with a wealth of biological information to analyze and predict. Fully sequenced genomes made us to have additional insights into the functional properties of the encoded proteins and made it possible to develop new tools and schemes for functional biology on a proteomic scale. Among those are the yeast two-hybrid system, mass spectrometry and microarray: the technology of choice to detect protein-protein interactions. These functional insights emerge as networks of interacting proteins, also known as "pathway informatics" or "interactomics". Without exception it is no longer possible to make advances in the signaling/regulatory pathway studies without integrating information technologies with experimental technologies. In this paper, we will introduce the databases of protein interaction worldwide and discuss several challenging issues regarding the actual implementation of databases.

Characterization of the Interaction of Sulfiredoxin (Srx1) with a Vacoular Protein $\alpha$-Mannosidase (Ams1) in Saccharomyces cerevisiae (설피리독신과 알파-만노시다제 간의 단백질 결합 특성에 관한 고찰)

  • Barando, Karen P.;Kim, Il-Han
    • The Journal of Natural Sciences
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    • v.17 no.1
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    • pp.13-29
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    • 2006
  • Most redox-active proteins have thiol-bearing cysteine residues that are sensitive to oxidation. Cysteine thiols oxidized to sulfenic acid are generally unstable, either forming a disulfide with a nearby thiol or being further oxidized to a stable sulfinic acid, which have been viewed as an irreversible protein modification. However, recent studies showed that cysteine residues of certain thiol peroxidases (Prxs) undergo reversible oxidation to sulfinic acid and the reduction reaction is catalyzed by sulfiredoxin (Srx1). Specific Cys residues of various other proteins are also oxidized to sulfinic acid ($Cys-So_2H$). Srxl is considered one of the oxidant proteins with a role in signaling through catalytic reduction of oxidative modification like in the reduction of glutathionylation, a post-translational, oxidative modification that occurs on numerous proteins. In this study, the role of sulfiredoxin in cellular processes, was investigated by studying its interaction with other proteins. Through the yeast two-hybrid system (Y2HS) technique, we have found that Ams1 is a potential and novel interacting protein partner of Srxl. $\alpha$-mannosidase (Ams1) is a resident vacuolar hydrolase which aids in recycling macromolecular components of the cell through hydrolysis of terminal, non-reducing $\alpha$-D-mannose residues. It forms an oligomer in the cytoplasm and under nutrient rich condition and is delivered to the vacuole by the Cytoplasm to Vacuole (Cvt) pathway. Aside from the role of Srxl as a catalyst in the reduction of cysteine sulfenic acid groups, it may play a completely new function in the cellular process as indicated by its interaction with Ams1 of the yeast Saccharomyces cerevisiae.

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Identification and Characterization of the Interaction between Heat-Shock Protein 90 and Phospholipase C-γ1

  • Kim, Su-Jeong;Kim, Myung-Jong;Kim, Yong;Si, Fu Chun;Ryu, Sung-Ho;Suh, Pann-Chill
    • BMB Reports
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    • v.33 no.2
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    • pp.97-102
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    • 2000
  • Phosphoinositide-specific phospholipase C-${\gamma}1$ (PLC-${\gamma}1$) is a pivotal mediator in the signal transduction cascades induced by many growth factors. Using a yeast two-hybrid system, heat-shock protein 90 (Hsp90) was identified as a PLC-${\gamma}1$-binding protein. A co-immunoprecipitation experiment, using anti-PLC-${\gamma}1$ antibody, demonstrated an in vivo interaction between Hsp90 and PLC-${\gamma}1$ in the NIH-3T3 cells. The interaction in NIH-3T3 was unaffected by the PDGF treatment, inducing phosphorylation and activation of PLC-${\gamma}1$. Direct interaction between Hsp90 and PLC-${\gamma}1$ was confirmed by in vitro binding experiments using purified Hsp90 and PLC-${\gamma}1$. Furthermore, Hsp90 increased the $PIP_2$-hydrolyzing activity of PLC-${\gamma}1$ up to 2-fold at $0.1{\mu}M$ in vitro. Taken together, we show for the first time, the interaction of PLC-${\gamma}1$ with Hsp90, both in vivo and in vitro. We suggest that Hsp90 may play a role in PLC-${\gamma}1$-mediated signal transduction.

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Construction of Recombinant DNA for Purification of the Gag-Pro Transframe Protein of Human T-cell Leukemia Virus Type I (HTLV-I) (Human T-cell Leukemia Virus Type I (HTLV-I) 의 Gag-Pro Transframe 단백질 정제를 위한 재조합 DNA 의 제작)

  • 남석현
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.466-471
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    • 1992
  • To determine the site at which -1 ribosomal frameshifting occurs within the gag-pro overlap of HTL V-I. DNA fragment corresponding to a portion of the gene overlap was cloned into a SP6 vector. The resultant plasmid harbors the hybrid gene consisting of a synthetic gene encoding 5 amino acids derived from chick prelysozyme including the initiator methionine plus 141 nucleotides of gag-pro overlapping region followed by Staphylococcus aurcus protein A gene fragment. In vitro transcription by SP6 RNA polymerase with this DNA template made an abundant amount of single species mRNA. Cell-free translation programmed with the RNA transcribed in vitro yielded a polypeptide of 21 kDal in size. which could be purified into homogeneity by IgG-Sepharose affinity chromatography. In vitro system described in this study must be useful for rapid purification and sequencing of the Gag-Pro transframe protein. allowing to determine the exact frameshift site on mRNA and to identify the tRNA involved in frameshifting event for the expression of pro gene.

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Nebulin C-terminus Interacts with NCBP51, a New Isoform of RING Finger Protein 125 (RNF125)

  • Kim, Ji-Hee;Kim, Hyun-Suk;Park, Eun-Ran;Choi, Jae-Kyoung;Lee, Yeong-Mi;Choi, Jun-Hyuk;Shin, Jung-Woog;Kim, Chong-Rak
    • Biomedical Science Letters
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    • v.13 no.1
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    • pp.1-10
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    • 2007
  • Nebulin, a giant modular protein from muscle, is thought to act as molecular ruler in sarcomere assembly. In skeletal muscle, the C-terminal ${\sim}50 kDa$ region of nebulin extends into the Z-line lattice. The most recent studies implicated highlighting its extensive isoform diversity and exciting reports revealed its expression in cardiac and non-muscle tissues containing brain. Also these novel findings are indicating that nebulin is actually a multifunctional filament system, perhaps playing roles in signal transduction, contractile regulation, and myofibril force generation, as well as other not yet defined functions. However the binding protein of nebulin and function in brain is still unknown. A novel binding partner of nebulin C-terminal region was identified by screening a human brain cDNA library using yeast two-hybrid system. Nebulin C-terminus binding protein 51 (NCBP51) was contained a RING-finger domain and identified a new isoform of RING finger protein 125 (RNF125). The interaction was confirmed using the GST pull-down assay. NCBP51 belongs to a family of the RING finger proteins and its function remains to be identified in brain. The role of nebulin and NCBP51 will be studied by loss-of-function using siRNA technique in brain.

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