• Title/Summary/Keyword: C1-2 fusion

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Analysis of the NS4 Region of Japanese Encephalitis virus K94P05 Isolated from Korea (일본뇌염 바이러스 국내분리주 K94P05의 NS4 부위 분석)

  • Kim, Eun-Jung;Nam, Jae-Hwan;Park, Yong-Kenun;Cho, Hae-Wol
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.197-207
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    • 1997
  • To investigate the NS4 region of JEV, NS4 cDNA of K94P05 (JEV strain isolated from Korea in 1994) was amplified by RT-PCR and analyzed by sequencing PCR product. Genomic size of NS4 was 1212bp and nucleotide sequence was compared with that of other JEV strains. Nucleotide homology between JaOAr582 and K94P05 was 91.1% and that between Beijing and K94P05 was 89.8%, respectively. But the nucleotide sequence of E region of JaOAr582 and K94P05 showed 97.0% homology and that of Beijing and K94P05 did 95.8% homology. NS4 protein was expressed as a form of fusion protein by a prokaryotic expression system. The induced fusion product showed a lower molecular weight than predicted size and remained insoluble. The NS4 protein might be cleavaged by E. coli protease. Concluding above results, high hydrophobicity of the NS4 protein supported the fact that this protein played a role as a membrane component and the poor nucleotide sequence conservativity among JEV strains suggested that this region might be important to adapt each viral growth environment.

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Improvement of L-Lysine Productivity by Using Cell Fusion and Immobilized System (세포융합과 고정화 시스템을 이용한 L-Lysine의 생산성 향상)

  • Ryu, Beung-Ho;Kim, Hye-Sung;Roh, Myung-Hoon;Park, Bob-Gyu;Chung, Jong-Soon;Bai, Ki-Chul
    • Korean Journal of Food Science and Technology
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    • v.21 no.1
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    • pp.154-163
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    • 1989
  • This studies were designed to improve the productivity of L-lysine by protoplast fusion and immobilized system of fusants using strains of Brevibacterium flavum ATCC 21528, Brevibacterium lactofermentum ATCC 21086 and Corynebacterium glutamicum 820. Mutants were isolated with concentration method of $300{\mu}g/ml$ penicillin-G after treatment of $250{\mu}g/ml$ N-methyl-N-nitro-N-nitrosoguanidine. B. flavum $37-2(Hos^-,\;Kan^r,\;AEC^r)$, B. lactofermentum $6-2(Ile^-,\;Val^-,\;Str^r,\;AEC^r)$ and C. glutamicum 57-5$(Met^-,\;Thr^-,\;Rif^r,\;AEC^r)$ were isolated from mutants. Protoplasts were induced by being incubated with $500{\mu}g/ml$ lysozyme of lysis solution for 6 hr and the ratio of protoplast formation and regeneration were ranging from 97-99% and 33-37%, respectively. Fusion frequencies of fusants of BBFL 21, BCFG 37 and BCLG 59 were shown in the range from $1.25{\times}10^{-6}\;to\;5.83{\times}10^{-7}$ under the optimum conditions. The fusant BBFL 21 showed the highest productivity of $411.1\;ng/ml{\cdot}hr$ L-lysine in the lysine productivity broth at $30^{\circ}C$ for 72hr. In the immobilization systems, fusant BBFL 21 was employed in various polymer matrices such as sodium alginate, polyacrylamide, agar and ${\alpha}-carrageena$. The immobilization of sodium alginate showed the highest productivity of $413\;ng/ml{\cdot}hr$ L-lysine in the batch system. Continuous fermentation of immobilization system by using tube fermentor was produced the highest productivity $416.7\;ng/ml{\cdot}hr $ L-lysine under optimum condition.

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Theoretical studies on the stabilization and diffusion behaviors of helium impurities in 6H-SiC by DFT calculations

  • Obaid Obaidullah;RuiXuan Zhao;XiangCao Li;ChuBin Wan;TingTing Sui;Xin Ju
    • Nuclear Engineering and Technology
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    • v.55 no.8
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    • pp.2879-2888
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    • 2023
  • In fusion environments, large scales of helium (He) atoms are produced by a radical transformation along with structural damage in structural materials, resulting in material swelling and degradation of physical properties. To understand its irradiation effects, this paper investigates the stability, electronic structure, energetics, charge density distribution, PDOS and TDOS, and diffusion processes of He impurities in 6HSiC materials. The formation energy indicates that a stable, favorable position for interstitial He is the HR site with the lowest energy of 2.40 eV. In terms of vacancy, the He atom initially prefers to substitute at pre-existing Si vacancy than C vacancy due to lower substitution energy. The minimum energy paths (MEPs) with migration energy barriers are also calculated for He impurity by interstitial and vacancy-mediated diffusion. Based on its calculated energy barriers, the most possible diffusion path includes the exchange of interstitial and vacancy sites with effective migration energies ranging from 0.101 eV to 1.0 eV. Our calculation provides a better understanding of the stabilization and diffusion behaviors of He impurities in 6H-SiC materials.

mCR1aa Medium를 이용한 소 복제수정란 배양 및 융합방법에 따른 발육율 비교

  • 김현주;양병철;임기순;오성종
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.63-63
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    • 2001
  • 본 실험은 소에서 체외수정란과 복제수정란을 modified CRlaa (mCRlaa) medium에서 각각 배양하였을 때 배반포단계까지 발육율을 비교하였다. 체외성숙은 TCM-199 medium에 10 %의 Fetal Bovine Serum (FBS)를 첨가한 배양액에서 실시하였고 이를 이용해 체외수정과 복제를 시행하였다. 체외성숙란과 복제수정란의 배반포까지 발육율은 각각 22.5 %, 19.4 %이었다. 복제수정란을 생산하기 위해서는 제핵을 실시한 난자내로 공여핵를 도입하는 과정이 필요한 데 할구보다 체세포를 이용하는 경우 융합율이 저하되는 것으로 보고되고 있다. 따라서 본 연구에서는 공여핵을 제공하는 체세포와 수핵란간에 융합을 기존에 많이 사용한 chamber와 비교해 Needle을 사용했을 때 복제수정란의 발육율간의 차이를 서로 비교하였다. Ear skin cell을 이용해 핵이식을 실시한 다음 Zimmerman cell fusion medium에서 융합하였다 융합된 핵이식란은 5 % $CO_2$, 5 % $O_2$, 90 % Na, 38.5$^{\circ}C$ 조건에서 배반포까지 배양하였다. Chamber와 Needle을 사용한 경우 융합율은 각각 46.1 %, 70.4 %, 분할율은 62.6 %, 76.4 %로 Needle의 경우가 유의적으로 높았다.

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Flavanone 3β-Hydroxylases from Rice: Key Enzymes for Favonol and Anthocyanin Biosynthesis

  • Kim, Jeong Ho;Lee, Yoon Jung;Kim, Bong Gyu;Lim, Yoongho;Ahn, Joong-Hoon
    • Molecules and Cells
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    • v.25 no.2
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    • pp.312-316
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    • 2008
  • Flavanone $3{\beta}$-hydroxylases (F3H) are key enzymes in the synthesis of flavonol and anthocyanin. In this study, three F3H cDNAs from Oryza sativa (OsF3H-1 ~3) were cloned by RT-PCR and expressed in E. coli as gluthatione S-transferase (GST) fusion proteins. The purified recombinant OsF3Hs used flavanone, naringenin and eriodictyol as substrates. The reaction products with naringen and eriodictyol were determined by nuclear magnetic resonance spectroscopy to be dihydrokaempferol and taxifolin, respectively. OsF3H-1 had the highest enzymatic activity whereas the overall expression of OsF3H-2 was highest in all tissues except seeds. Flavanone $3{\beta}$-hydroxylase could be a useful target for flavonoid metabolic engineering in rice.

Genetic Recombination by Protoplast Fusion of Cellulomonas sp CS 1-1 (원형질체 융합법에 의한 cellulomonas sp. CS 1-1의 유전자 재조합)

  • 김병홍;이학주
    • Korean Journal of Microbiology
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    • v.23 no.4
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    • pp.309-314
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    • 1985
  • Autoxotrophic mutants of a cellulolytic baterium Cellulomonas sp. CS 1-1 were grown at $30^{\circ}C$ for 6hr using a complete medium containing 0.5M sucrose and for another 90 min after addition of 0.3 U/ml penicillin G, and were protoplasted by 0.2mg/ml lysozyme for 2hr. Prototrophic recombinants were obtained at the rates of $10^{-3}$ to $10^{-5}$by fusing the protoplasts in the presence of 40% polyethyleneglycol3350. Nystatin could be used to eliminate fungal contamination during the regeneration of the plotaplasts.

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Characterization of the Methylation Status of Pax7 and Myogenic Regulator Factors in Cell Myogenic Differentiation

  • Chao, Zhe;Zheng, Xin-Li;Sun, Rui-Ping;Liu, Hai-Long;Huang, Li-Li;Cao, Zong-Xi;Deng, Chang-Yan;Wang, Feng
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.7
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    • pp.1037-1043
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    • 2016
  • Epigenetic processes in the development of skeletal muscle have been appreciated for over a decade. DNA methylation is a major epigenetic modification important for regulating gene expression and suppressing spurious transcription. Up to now, the importance of epigenetic marks in the regulation of Pax7 and myogenic regulatory factors (MRFs) expression is far less explored. In the present study, semi-quantitative the real-time polymerase chain reaction (RT-PCR) analyses showed MyoD and Myf5 were expressed in activated and quiescent C2C12 cells. MyoG was expressed in a later stage of myogenesis. Pax7 was weakly expressed in differentiated C2C12 cells. To further understand the regulation of expression of these genes, the DNA methylation status of Pax7, MyoD, and Myf5 was determined by bisulfite sequencing PCR. During the C2C12 myoblasts fusion process, the changes of promoter and exon 1 methylation of Pax7, MyoD, and Myf5 genes were observed. In addition, an inverse relationship of low methylation and high expression was found. These results suggest that DNA methylation may be an important mechanism regulating Pax7 and MRFs transcription in cell myogenic differentiation.

The Effect of Polymer Blending and Extension Conditions on the Properties of Separator Prepared by Wet Process for Li-ion Secondary Battery (고분자 블렌딩 및 연신조건이 리튬 이온전지용 습식 Separator의 물성에 미치는 영향)

  • 문성인;손영수;김순식;김진열
    • Polymer(Korea)
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    • v.26 no.1
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    • pp.45-52
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    • 2002
  • The separator made from the blends of high density polyethylene (HDPE) and ultrahigh molecular weight polyethylene (UHMWPE) was prepared by wet processing to use as Li-ion secondary battery. We investigated effects of the blending of the polymers and the film extension on the mechanical properties of the separator. The mechanical strength of separator increased with increasing molecular weights and contents of UHMWPE, for instance about $1000 kg/\textrm{cm}^2$ with the five times extended film of 6 wt% UHMWPE. The pores of the separator were very uniform with the size of 0.1~$0.12\mu\textrm{m}$. The shut-down characteristic quickly increased at around $130^{\circ}C$ and the fusion temperature was $160^{\circ}C$, so it could be applied to the lithium ion secondary battery.

ZNF435, a Novel Human SCAN-containing Zinc Finger Protein, Inhibits AP-1-mediated Transcriptional Activation

  • Gu, Xing;Zheng, Mei;Fei, Xiangwei;Yang, Zhenxing;Li, Fan;Ji, Chaoneng;Xie, Yi;Mao, Yumin
    • Molecules and Cells
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    • v.23 no.3
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    • pp.316-322
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    • 2007
  • Zinc finger transcription factor genes are a significant fraction of the genes in the vertebrate genome. Here we report the isolation and characterization of a human zinc finger-containing gene, ZNF435, from a fetal brain cDNA library. ZNF435 cDNA is 1290 base pairs in length and contains an open reading frame encoding 349 amino acids with four C2H2-type zinc fingers at its carboxyl terminus and a SCAN motif at its amino terminus. RT-PCR results showed that ZNF435 was expressed in all tested tissues. A ZNF435-GFP fusion protein was located in the nucleus and the four zinc fingers acted as nuclear localization signals (NLSs). ZNF435 was found to be capable of homo-association, and this effect was independent of its zinc fingers. Furthermore, ZNF435 proved to be a transcription repressor as its overexpression in AD293 cells inhibited the transcriptional activities of AP-1.

Study on the Thermal Storage Characteristics of a Multi-capsule type LTES System -Analysis for Heat Charging and Discharging Process for Water Flow- (다관형 잠열축열장치의 축열특성연구 -물을 매체로 한 축열 및 방열과정 분석-)

  • Kim, Y.B.
    • Journal of Biosystems Engineering
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    • v.19 no.1
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    • pp.62-69
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    • 1994
  • This study was designed to seek information on the heat charging and discharging characteristics of a multi-capsule type LTES(Latent Heat of Fusion Thermal Energy Storage) system, and especially prediction equation of outlet water temperature from the system. During heat charging process, the water temperature in the LTES tank increased very slowly in comparison with a predicted one and was kept near the melting point of the PCM for about 25 minutes. During heat discharging process, the latent heat discharging period of the outlet water temperature became longer as the inlet water temperature became higher and/or mass flow rate became lower. The dimensionless temperature of the outlet water was predicted by linking three equations of ${\theta}=1.1Exp(-{\tau}/0.82)$, ${\theta}=-0.06{\tau}+0.3$, ${\theta}=0.8Exp(-{\tau}/1.4)$ ($r^2{\leq}0.88$) depending on discharging period regardless of mass flow rates on the case of the inlet water temperature at $21.5^{\circ}C$.

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