• Title/Summary/Keyword: heavy chain

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PCR Specific Primer for the Detection of Vibrio tapetis (Vibrio tapetis의 검출을 위한 PCR specific primer의 제작)

  • Kim, Yeong-Jin;Lee, Sun-Yi;Cho, Hyo-Jin;You, Sun-Nyung;Kim, Cheol-Min;Choi, Yong-Lark;Park, Byoung-Keun;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.17 no.1 s.81
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    • pp.162-165
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    • 2007
  • Brown Ring Disease (BRD) is a bacterial disease caused by Vibrio tapetis which affects cultured clam Ruditapes philippinarum and causes heavy economic losses on Atlantic coasts of france, Spain and England. In this study, to evaluate the effective detection of the pathogen, specific primer set based on 16S ribosomal RNA (rRNA) sequences designed for rapid detection of V. tapetis. Polymerase chain reaction (PCR) with this primer set produced the specific band for each V. tapetis. The length of PCR product using designed primer set of Vbts-F and Vbts-R was about 400 bp. Therefore, these primers will be provided with a basic tool for rapid detection of V. tapetis in the various cases such as examination of imported aquatic products, diagnosis of aquatic organisms, and etc.

The Efficiency of RNA Interference in Bursaphelenchus xylophilus

  • Park, Jung-Eun;Lee, Kyong Yun;Lee, Se-Jin;Oh, Wan-Suk;Jeong, Pan-Young;Woo, Taeha;Kim, Chang-Bae;Paik, Young-Ki;Koo, Hyeon-Sook
    • Molecules and Cells
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    • v.26 no.1
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    • pp.81-86
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    • 2008
  • RNA interference (RNAi) was performed on several essential genes in the pinewood nematode Bursaphelenchus xylophilus, which causes pine wilt disease. Double-stranded RNA (dsRNA) was delivered to larvae or adult worms by soaking, electroporation, or microinjection. Soaking and electroporation of L2-L3 stage worms in solutions containing dsRNA for essential genes induced over 25% lethality after 5 days, and gene-specific phenotypes were observed. This lethality agreed with significant reductions of the targeted transcripts, as assayed by reverse-transcription coupled with real time PCR. Microinjection was the most efficient route as measured by the hatching rate of F1 embryos, which was reduced by 46%. When adult worms were soaked in dsRNA, lethality was induced in the F1 larvae, revealing the persistence of knockdown phenotypes. The penetrance of the RNAi phenotypes for essential genes was relatively low but consistent, indicating that RNAi should be useful for studying the in vivo functions of B. xylophilus gene products.

Modification of Rubbers through Chemical Reactions including Controlled/"living" Radical Polymerization Techniques (리빙라디칼 중합법을 포함한 화학적 방법에 의한 고무의 개질)

  • Joo, Sang-Il;Cho, Hyun-Chul;Lee, Seong-Hoon;Hong, Sung-Chul
    • Elastomers and Composites
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    • v.44 no.2
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    • pp.122-133
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    • 2009
  • Rubbers, such as natural rubber, polybutadiene, styrene-butadiene rubber, nitrile-butadiene rubber, chlorinated rubber and EPDM, have been continuously improved in response to a heavy demand and a new property requirement from industry. One of the best ways to realize the improvement is the modification of rubbers through chemical reactions, which produce materials with novel properties. In this review, chemical modification reactions of rubbers that contain carbon-carbon double bond units either in their main backbone or as a side group were briefly summarized. The chemical reactions introduce functional groups or functional polymer chains to polymer backbone, which transform a classical rubber to a highly functional material. Especially, we focused on a controlled/"living" radical polymerization techniques, with which a revolutionary broadening of the spectrum of the materials with well defined molecular weight, molecular weight distribution, chain end-functionality and architectures become possible.

Effects of Bromelain and Double Emulsion on the Physicochemical Properties of Pork Loin

  • Shin, Hyerin;Kim, Hyo Tae;Choi, Mi-Jung;Ko, Eun-Young
    • Food Science of Animal Resources
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    • v.39 no.6
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    • pp.888-902
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    • 2019
  • Our aim was to investigate the effects of bromelain embedded in double emulsion (DE) on physicochemical properties of pork loin. We evaluated DE characteristics such as size, zeta potential, and microscopy after fabrication. We marinated meat with distilled water (DW), 1% (w/v) bromelain solution, blank DE, and 1% (w/v) bromelain loaded in double emulsion (DE E) for 0, 24, 48, and 72 h at 4℃, and prepared raw meat for control. The marinated samples were assessed for color, water holding capacity, cooking loss, moisture content, pH, protein solubility, Warner-Bratzler shear force (WBSF) and gel electrophoresis. The droplet size of 1% (w/v) bromelain embedded in DE was increased compared with blank DE (p<0.05) and values of zeta potential decreased. The increase in lightness and color difference range of the DE-treated group was lower than that of the DW-treated group (p<0.05). Moreover, treatment by immersion in 1% (w/v) DE E resulted in the highest water holding capacity values (p<0.05) and lower cooking loss values than water base treatment (p<0.05). Results for myofibrillar protein solubility and WBSF showed a similar trend. 1% (w/v) DE E showed degradation of myosin heavy chain after 48 h in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Thus, bromelain-loaded DE is useful for controlling and handling enzyme activity in food industry.

Proteomic Analysis of Differentially Expressed Proteins in Bovine Endometrium with Endometritis

  • Choe, Chang-Yong;Park, Jeong-Won;Kim, Eun-Suk;Lee, Sung-Gyu;Park, Sun-Young;Lee, Jeong-Soon;Cho, Myung-Je;Kang, Kee-Ryeon;Han, Jae-Hee;Kang, Da-Won
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.4
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    • pp.205-212
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    • 2010
  • Endometritis is one of the primary reasons for reproductive failure. In order to investigate endometritis-associated marker proteins, proteomic analysis was performed on bovine endometrium with endometritis. In bovine endometritis, desmin, $\alpha$-actin-2, heat-shock protein (HSP) 27, peroxiredoxin-6, luteinizing hormone receptor isoform 1, collectin-43 precursor, deoxyribonuclease-I (DNase-I), and MHC class I heavy chain (MHC-Ih) were up-regulated. In contrast, transferrin, interleukin-2 precursor, hemoglobin $\beta$ subunit, and potassium channel tetramerisation domaincontaining 11 (KCTD11) were down-regulated in comparison to normal endometrium. The proteomic results were validated by semiquantitative-PCR and immunoblot analysis. The mRNA levels of desmin, transferrin, $\alpha$-actin-2, HSP27, KCTD11, and MHC-Ih were up-regulated by over 1.5-fold, and showed a pattern similar to their proteomic profiles. Desmin and $\alpha$-actin-2 protein showed positive correlations between proteomic analysis and immunoblot analysis. These results suggest that desmin and $\alpha$-actin-2 may play important roles in endometritis-related function, and could be useful markers for the diagnosis of bovine endometritis.

JPH203, a selective L-type amino acid transporter 1 inhibitor, induces mitochondria-dependent apoptosis in Saos2 human osteosarcoma cells

  • Choi, Dae Woo;Kim, Do Kyung;Kanai, Yoshikatsu;Wempe, Michael F.;Endou, Hitoshi;Kim, Jong-Keun
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.6
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    • pp.599-607
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    • 2017
  • Most normal cells express L-type amino acid transporter 2 (LAT2). However, L-type amino acid transporter 1 (LAT1) is highly expressed in many tumor cells and presumed to support their increased growth and proliferation. This study examined the effects of JPH203, a selective LAT1 inhibitor, on cell growth and its mechanism for cell death in Saos2 human osteosarcoma cells. FOB human osteoblastic cells and Saos2 cells expressed LAT1 and LAT2 together with their associating protein 4F2 heavy chain, but the expression of LAT2 in the Saos2 cells was especially weak. JPH203 and BCH, a non-selective L-type amino acid transporter inhibitor, potently inhibited L-leucine uptake in Saos2 cells. As expected, the intrinsic ability of JPH203 to inhibit L-leucine uptake was far more efficient than that of BCH in Saos2 cells. Likewise, JPH203 and BCH inhibited Saos2 cell growth with JPH203 being superior to BCH in this regard. Furthermore, JPH203 increased apoptosis rates and formed DNA ladder in Saos2 cells. Moreover, JPH203 activated the mitochondria-dependent apoptotic signaling pathway by upregulating pro-apoptotic factors, such as Bad, Bax, and Bak, and the active form of caspase-9, and downregulating anti-apoptotic factors, such as Bcl-2 and Bcl-xL. These results suggest that the inhibition of LAT1 activity via JPH203, which may act as a potential novel anti-cancer agent, leads to apoptosis mediated by the mitochondria-dependent intrinsic apoptotic signaling pathway by inducing the intracellular depletion of neutral amino acids essential for cell growth in Saos2 human osteosarcoma cells.

Influence of NaCl and pH on Hydrolysis of Chicken Myofibrillar Proteins by Leukocyte Lysosomal Proteinases (Leucocyte lysosomal proteinase에 의한 닭의 근섬유(筋纖維) 단백질(蛋白質) 분해(分解)에 미치는 NaCl과 pH의 영향(影響))

  • Shinlee, Seung-Yee;Rhee, Chong-Ouk
    • Korean Journal of Food Science and Technology
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    • v.22 no.5
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    • pp.569-574
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    • 1990
  • The influence of NaCl and pH on degradation of chicken breast muscle myofibrillar proteins by porcine leukocyte lysosomal proteinases was investigated. The degradation reactions were carried out at $38^{\circ}C$ for 24hours under different conditions. The degradation of myofibrillar proteins by leukocyte lysosomal enzymes at various pH values was limited to partial hydrolysis. Reactions at higher pH values resulted in lower molecular weight degradation products while reactions at lower pH resulted in higher molecular weight degradation products. When NaCl was added into the reaction mixture, enzyme activities of degradation were increased at all pH values studied, as evidenced by NPN-analysis and SDS-PAGE. More severe degradation was observed with higher salt concentration. The concentration of 0.5M NaCl in the reaction mixture gave more degradation of myosin heavy chain by enzyme than that of 0.1M NaCl.

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The effect of rod domain A148V mutation of neurofilament light chain on filament formation

  • Lee, In-Bum;Kim, Sung-Kuk;Chung, Sang-Hee;Kim, Ho;Kwon, Taeg-Kyu;Min, Do-Sik;Chang, Jong-Soo
    • BMB Reports
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    • v.41 no.12
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    • pp.868-874
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    • 2008
  • Neurofilaments (NFs) are neuronal intermediate filaments composed of light (NF-L), middle (NF-M), and heavy (NF-H) subunits. NF-L self-assembles into a "core" filament with which NF-M or NF-H co-assembles to form the neuronal intermediate filament. Recent reports show that point mutations of the NF-L gene result in Charcot-Marie-Tooth disease (CMT). However, the most recently described rod domain mutant of human NF-L (A148V) has not been characterized in cellular level. We cloned human NF-L and used it to engineer the A148V. In phenotypic analysis using SW13 cells, A148V mutation completely abolished filament formation despite of presence of NF-M. Moreover, A148V mutation reduced the levels of in vitro self-assembly using GST-NF-L (H/R) fusion protein whereas control (A296T) mutant did not affect the filament formation. These results suggest that alanine at position 148 is essentially required for NF-L self-assembly leading to subsequent filament formation in neuronal cells.

Two Kinesins from Arabidopsis, KatB and KatC, Have a Second Microtubule-binding Site in the Tail Domain

  • Jiang, Shiling;Li, Ming;Xu, Tao;Ren, Dongtao;Liu, Guoqin
    • BMB Reports
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    • v.40 no.1
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    • pp.44-52
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    • 2007
  • Kinesins, as a kind of microtubule-based motor proteins, have a conserved microtubule-binding site in their motor domain. Here we report that two homologous kinesins in Arabidopsis thaliana, KatB and KatC, contain a second microtubule-binding site in their tail domains. The prokaryotic-expressed N-terminal tail domain of the KatC heavy chain can bind to microtubules in an ATP-insensitive manner. To identify the precise region responsible for the binding, a serious of truncated KatC cDNAs encoding KatC N-terminal regions in different lengths, KatC1-128, KatC1-86, KatC1-73 and KatC1-63, fused to Histidine-tags, were expressed in E. coli and affinity-purified. Microtubule cosedimentation assays show that the site at amino acid residues 74-86 in KatC is important for microtubule-binding. By similarity, we obtained three different lengths of KatB N-terminal regions, KatB1-384, KatB1-77, and KatB1-63, and analyzed their microtubule-binding ability. Cosedimentation assays indicate that the KatB tail domain can also bind to microtubules at the same site as and in a similar manner to KatC. Fluorescence microscopic observations show that the microtubule-binding site at the tail domain of KatB or KatC can induce microtubules bundling only when the stalk domain is present. Through pull-down assays, we show that KatB1-385 and KatC1-394 are able to interact specifically with themselves and with each other in vitro. These findings are significant for identifying a previously uncharacterized microtubule-binding site in the two kinesin proteins, KatB and KatC, and the functional relations between them.

Performance Evaluation of an Adaptive Collision Avoidance Algorithm for IEEE 802.15.4 (IEEE 802.15.4에서 적응적 충돌 회피 알고리즘의 성능분석)

  • Noh, Ki-Chul;Lee, Seung-Yeon;Shin, Youn-Soon;Ahn, Jong-Suk;Lee, Kang-Woo
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.36 no.3A
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    • pp.267-277
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    • 2011
  • Like other wireless network protocols, IEEE 802.15.4 adopts CA (Collision Avoidance)algorithm to avoid the early collision of a new packet by randomizing its first transmission time rather than its immediate delivery. The traditional CA scheme of IEEE 802.15.4, however, selects the random access time from the predetermined range without considering the degree of current congestion. It probably causes either a long delay to settle in the suitable range for the current network load or frequent clashes especially during the long lasting heavy traffic period. This paper proposes an ACA(Adaptive Collision Avoidance) algorithm adapting the initial backoff range to the undergoing collision variations. It also introduces a mathematical model to predict the performance of ACA algorithm added to IEEE 802.15.4. With only small deviations from corresponding simulations, our analytical model shows that ACA technique can improve the throughput of IEEE 802.15.4 by up-to 41% maximally in addition to shortening the packet delays.