• Title/Summary/Keyword: heavy chain

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Construction of fluorescent red silk using fibroin H-chain expression system (누에 형질전환에 의한 견사선에서의 적색형광단백질 발현)

  • Kim, Sung Wan;Yun, Eun Young;Choi, Kwang-Ho;Kim, Seong Ryul;Park, Seung Won;Kang, Seok Woo;Kwon, O-Yu;Goo, Tae Won
    • Journal of Sericultural and Entomological Science
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    • v.50 no.2
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    • pp.87-92
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    • 2012
  • We constructed the fibroin H-chain expression system to produce Discosoma sp. red fluorescent protein variant2 (DsRed2) in transgenic silkworm cocoon. Fluorescent cocoon could be made by fusing DsRed2 cDNA to the heavy chain gene and injecting it into a silkworm. The DsRed2 fusion protein, each with N- and C-terminal sequences of the fibroin H-chain, was designed to be secreted into the lumen of the posterior silk glands. The expression of the DsRed2/H-chain fusion gene was regulated by the fibroin H-chain promoter. The use of the 3xP3-driven EGFP cDNA as a marker allowed us to rapidly distinguish transgenic silkworms. The EGFP fluorescence became visible in the ocelli and in the central and peripheral nervous system on the seventh day of embryonic development. A mixture of the donor and helper vector was micro-injected into 1,020 Kumokjam, bivoltin silkworm eggs. We obtained 6 broods. The cocoon was displayed strong red fluorescence, proving that the fusion protein was present in the cocoon. Accordingly, we suggest that the DsRed2 fluorescence silk will enable the production of novel biomaterial based on the transgenic silk.

Rab Effector EHBP1L1 Associates with the Tetratricopeptide Repeat Domain of Kinesin Light Chain 1 (Kinesin Light Chain 1 (KLC1)의 Tetratricopeptide Repeat (TPR) 도메인과 Rab effector, EHBP1L1의 결합)

  • Jeong, Young Joo;Park, Sung Woo;Kim, Sang-Jin;Kim, Mooseong;Urm, Sang-Hwa;Lee, Jung Goo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.30 no.1
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    • pp.10-17
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    • 2020
  • Kinesin-1 is microtubule-dependent plus-end direct molecular motor protein essential for intracellular transport. It is a member of the kinesin superfamily proteins (KIFs) which transport cargo, including organelles, vesicles, neurotransmitter receptors, cell-signaling molecules, and protein complexes through interaction between its light chain subunit and the cargo. Kinesin light chain 1 (KLC1) is a non-motor subunit that associates with the kinesin heavy chain (KHC). Although KLC1 interacts with many different adaptor proteins and scaffolding proteins, its binding proteins have not yet been fully identified. We used the yeast two-hybrid assay to identify proteins that interact with the tetratricopeptide repeat (TPR) domain of KLC1, and found an interaction between KLC1 and EH domain-binding protein 1 like 1 (EHBP1L1). EHBP1L1 bound to the region containing all six TPR repeats of KLC1 and did not interact with KIF5B (a motor protein of kinesin 1) or KIF3A (a motor protein of kinesin 2) in the yeast two-hybrid assay. The carboxyl-terminus of the coiled-coil domain of EHBP1L1 is essential for interaction with KLC1. However, another EHBP1L1 isoform, EHBP1, did not interact with KLC1 in the yeast two-hybrid assay. KLC1 interacted with GST-EHBP1L1 and its coiled-coil domain but not with GST only. When co-expressed in HEK-293T cells, EHBP1L1 co-localized with KLC1 and co-immunoprecipitated with KLC1 and KIF5B but not KIF3A. These results suggest that kinesin 1 motor protein may transport EHBP1L1-associated cargo in cells.

Affinity Maturation of an Anti-Hepatitis B Virus PreS1 Humanized Antibody by Phage Display

  • Yang, Gi-Hyeok;Yoon, Sun-Ok;Jang, Myung-Hee;Hong, Hyo-Jeong
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.528-533
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    • 2007
  • In a previous study we generated an anti-Hepatitis B Virus (HBV) preS1 humanized antibody (HzKR127) that showed in vivo HBV-neutralizing activity in chimpanzees. However, the antigen-binding affinity of the humanized antibody may not be sufficient for clinical use and thus affinity maturation is required for better therapeutic efficacy. In this study, phage display technique was employed to increase the affinity of HzKR127. All six amino acid residues (Glu95-Tyr96-Asp97-Glu98-Ala99-Tyr100) in the heavy (H) chain complementary-determining region 3 (HCDR3) of HzKR127 were randomized and phage-displayed single chain Fv (scFv) library was constructed. After three rounds of panning, 12 different clones exhibiting higher antigen-binding activity than the wild type ScFv were selected and their antigen-binding specificity for the preS1 confirmed. Subsequently, five ScFv clones were converted to whole IgG and subjected to affinity determination. The results showed that two clones (B3 and A19) exhibited an approximately 6 fold higher affinities than that of HzKR127. The affinity-matured humanized antibodies may be useful in anti-HBV immunotherapy.

The Effects of Astragali Radix Extracts on Mitochondrial Function in C2C12 Myotubes (C2C12 골격근세포에서 황기의 미토콘드리아 조절 작용)

  • Song, Miyoung
    • Journal of Korean Medicine for Obesity Research
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    • v.14 no.2
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    • pp.55-62
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    • 2014
  • Objective: The prevalence of metabolic syndrome and type 2 diabetes is increasing worldwide. Mitochondrial dysfunction is known to be involved in insulin resistance and obesity, researches have been increasing highly. Astragali Radix extract (ARE) or its main components have been shown to perform comparably to insulin by significantly reducing blood glucose levels in animal models however, the influence on mitochondrial dysfunction are not well understood. Methods: ARE (0.2, 0.5 and 1.0 mg/ml) or metformin (2.5 mM) were treated in C2C12 after 6 day-differentiation. The expressions of adenosine monophosphate (AMP)-activated protein kinase (AMPK) and phosphorylation AMPK, peroxisome proliferators-activated receptror ${\gamma}$ coactivator $1{\alpha}$ ($PGC1{\alpha}$), nuclear respiratory factors 1 (NRF1), mitochondrial transcription factor (Tfam) and myosin heavy chain were detected with western blotting or polymerase chain reaction analysis. The morphological changes were also investigated. Results: ARE dose dependently increased phosphorylation of AMPK and respectively activated mRNA expressions of $PGC1{\alpha}$, NRF1 and Tfam which are mitochondrial biogenesis regulators. Furthermore, there were some morphologic differences of differentiated cells between ARE treatment and control. Conclusions: This study suggests that ARE has the potential to increase muscle mitochondrial function by activating AMPK and $PGC1{\alpha}$.

X-ray Diffraction Studies of Poly(aryl ether ether ketone) Fibers with Different Degrees of Crystallinity and Orientation

  • Karacan Ismail
    • Fibers and Polymers
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    • v.6 no.3
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    • pp.206-218
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    • 2005
  • Structural studies of series of 'as spun' and drawn PEEK fibers have been carried out using X-ray diffraction and optical microscopy techniques. The analysis of results suggest that fibers produced at a constant draw ratio with increasing draw temperatures show enhanced orientation and crystalline behaviour. The resolved equatorial and meridional traces provide additional structural parameters in terms of crystallinity, crystallite size, and crystallite thickness. It is concluded that drawing at a temperature below $T_g(i.e.,\;144^{\circ}C)$ results in poorly oriented non-crystalline materials, whereas drawing above $T_g$ results in highly oriented semi crystalline materials. Additional drawing proved to increase the overall orientation with slight improvements in lateral order of the chain molecules. Quantitative analysis revealed that the crystallite size increases with increasing drawing temperature. The results also revealed the increased crystallite size upon additional drawing. Crystalline orientation parameter, $_c$, suggests almost perfect orientation. In all cases, the amorphous orientation is found to be lower than the overall orientation parameter obtained from the optical birefringence. As a result of additional drawing, crystalline orientation was found to increase slightly but the increase in the orientation of non-crystalline material was found to be substantial. An average crystalline density was determined from the orthorhombic unit cell dimensions. It was found to vary as a result of processing conditions. It was also found that the value of the maximum birefringence shows heavy dependence on the chain conformation.

Infection with Citrobacter rodentium in μMT Knockout Mice

  • Jo, Minjeong;Hwang, Soonjae;Rhee, Ki-Jong
    • Biomedical Science Letters
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    • v.24 no.1
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    • pp.1-8
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    • 2018
  • ${\mu}MT$ knockout mice are genetically deficient in the transmembrane domain of mu chain of the immunoglobulin M (IgM) heavy chain, resulting in the absence of mature B cells. ${\mu}MT$ knockout mice is an in vivo model system used to clarify the role of B cells in various diseases. Enteropathogenic Escherichia coli (EPEC) induces acute and chronic diarrheal disease, especially in children of developing countries. The formation of attaching and effacing (A/E) lesion is a prominent pathogenic factor in the intestinal epithelium of EPEC infection. The A/E lesion is modulated by genes located on the pathogenic island locus of enterocyte effacement (LEE) which encode a type III secretion system (T3SS) and A/E lesion-related effector proteins. Citrobacter rodentium is a murine pathogen utilized in studying the pathogenic mechanisms of EPEC in human infections. Citrobacter rodentium produce A/E lesion to attach to intestinal epithelium, thus providing a murine model pathogen to study EPEC. Several studies have investigated the pathogenesis of Citrobacter rodentium in the ${\mu}MT$ knockout mice. In this review, we introduce the ${\mu}MT$ murine model in the context of C. rodentium pathogenesis and describe in detail the role of B cells and antibodies in this disease.

A Model for Analyzing the Performance of Wireless Multi-Hop Networks using a Contention-based CSMA/CA Strategy

  • Sheikh, Sajid M.;Wolhuter, Riaan;Engelbrecht, Herman A.
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.11 no.5
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    • pp.2499-2522
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    • 2017
  • Multi-hop networks are a low-setup-cost solution for enlarging an area of network coverage through multi-hop routing. Carrier sense multiple access with collision avoidance (CSMA/CA) is frequently used in multi-hop networks. Multi-hop networks face multiple problems, such as a rise in contention for the medium, and packet loss under heavy-load, saturated conditions, which consumes more bandwidth due to re-transmissions. The number of re-transmissions carried out in a multi-hop network plays a major role in the achievable quality of service (QoS). This paper presents a statistical, analytical model for the end-to-end delay of contention-based medium access control (MAC) strategies. These strategies schedule a packet before performing the back-off contention for both differentiated heterogeneous data and homogeneous data under saturation conditions. The analytical model is an application of Markov chain theory and queuing theory. The M/M/1 model is used to derive access queue waiting times, and an absorbing Markov chain is used to determine the expected number of re-transmissions in a multi-hop scenario. This is then used to calculate the expected end-to-end delay. The prediction by the proposed model is compared to the simulation results, and shows close correlation for the different test cases with different arrival rates.

Extreme Mooring Analysis of Turret Moored LNG-FSRU (터렛 계류된 LNG-FSRU의 극한 계류 해석)

  • Lee, Min-Kyeong;Jung, Kwang-Hyo;Park, Sung-Boo;Yu, Byeong-Seok;Chung, Yun-Suk
    • Journal of the Society of Naval Architects of Korea
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    • v.53 no.5
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    • pp.435-446
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    • 2016
  • In this study, hydrodynamic and mooring analysis for LNG FSRU moored by an internal turret with 9 mooring lines are numerically performed using commercial softwares, Hydrostar and Ariane. Met-ocean combinations for screening method are taken from wave governed condition(BV Rule Note NR 493) with relative heading between wave and wind between −45° and +45° and relative heading between wind and current between −30° and +30°. Extreme mooring analysis and sensitivity analysis are performed for intact and damaged (=one line missing) conditions and the parameters for sensitivity analysis are wave peak period, peak enhancement factor and line pretension. In the viewpoint of the design tension in mooring line, chain diameter is designed to satisfy safety factor for each conditions. As the chain diameter is increased from 152mm to 171mm, the designtension is reduced while the minimum breaking load is increased.

Prediction of extreme rainfall with a generalized extreme value distribution (일반화 극단 분포를 이용한 강우량 예측)

  • Sung, Yong Kyu;Sohn, Joong K.
    • Journal of the Korean Data and Information Science Society
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    • v.24 no.4
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    • pp.857-865
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    • 2013
  • Extreme rainfall causes heavy losses in human life and properties. Hence many works have been done to predict extreme rainfall by using extreme value distributions. In this study, we use a generalized extreme value distribution to derive the posterior predictive density with hierarchical Bayesian approach based on the data of Seoul area from 1973 to 2010. It becomes clear that the probability of the extreme rainfall is increasing for last 20 years in Seoul area and the model proposed works relatively well for both point prediction and predictive interval approach.

The Yin and Yang of RNA surveillance in B lymphocytes and antibody-secreting plasma cells

  • Lambert, Jean-Marie;Srour, Nivine;Delpy, Laurent
    • BMB Reports
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    • v.52 no.12
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    • pp.671-678
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    • 2019
  • The random V(D)J recombination process ensures the diversity of the primary immunoglobulin (Ig) repertoire. In two thirds of cases, imprecise recombination between variable (V), diversity (D), and joining (J) segments induces a frameshift in the open reading frame that leads to the appearance of premature termination codons (PTCs). Thus, many B lineage cells harbour biallelic V(D)J-rearrangements of Ig heavy or light chain genes, with a productively-recombined allele encoding the functional Ig chain and a nonproductive allele potentially encoding truncated Ig polypeptides. Since the pattern of Ig gene expression is mostly biallelic, transcription initiated from nonproductive Ig alleles generates considerable amounts of primary transcripts with out-of-frame V(D)J junctions. How RNA surveillance pathways cooperate to control the noise from nonproductive Ig genes will be discussed in this review, focusing on the benefits of nonsense- mediated mRNA decay (NMD) activation during B-cell development and detrimental effects of nonsense-associated altered splicing (NAS) in terminally differentiated plasma cells.