• Title/Summary/Keyword: MBP-E2

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Structure-Function Analysis of DNA Binding Domain of the Yeast ABF1 Protein (효모 ABF1 단백질의 DNA Binding 부위에 대한 구조 기능 연구)

  • Cho, Gi-Nam;Lee, Sang-Kyung;Kim, Hong-Tae;Kim, Ji-Young;Rho, Hyune-Mo;Jung, Gu-Hung
    • Korean Journal of Microbiology
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    • v.32 no.2
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    • pp.102-108
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    • 1994
  • Autonomously replicating sequence Binding Factor 1(ABF1) is a DNA-binding protein that specifically recognizes the $RTCRYN_5ACG$ at many sites in the yeast genome including the promoter element, mating-type silencer and ARS. To express the intact full-length ABF1 gene in E. coli, the ABF1 gene has been cloned into pMAL-c2 and His-61, Leu-353 and Leu-360 were substituted with other amino acid. ABF1 fusion proteins of wild type ABF1 and H61A, L353R and L360R nutants were purified by amylose resin affinity chromatography. Fusion protein of MBP and ABF1 was digested by Factor Xa and Characterized by gel retardation assay and complementation test. As aresult, we suggested that other DNA binding motif except atypical inc-finger motif is in the middle region of ABF1.

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LDPC Decoder for WiMAX/WLAN using Improved Normalized Min-Sum Algorithm (개선된 정규화 최소합 알고리듬을 적용한 WiMAX/WLAN용 LDPC 복호기)

  • Seo, Jin-Ho;Shin, Kyung-Wook
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.18 no.4
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    • pp.876-884
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    • 2014
  • A hardware design of LDPC decoder which is based on the improved normalized min-sum(INMS) decoding algorithm is described in this paper. The designed LDPC decoder supports 19 block lengths(576~2304) and 6 code rates(1/2, 2/3A, 2/3B, 3/4A, 3/4B, 5/6) of IEEE 802.16e mobile WiMAX standard and 3 block lengths(648, 1296, 1944) and 4 code rates(1/2, 2/3, 3/4, 5/6) of IEEE 802.11n WLAN standard. The decoding function unit(DFU) which is a main arithmetic block is implemented using sign-magnitude(SM) arithmetic and INMS decoding algorithm to optimize hardware complexity and decoding performance. The LDPC decoder synthesized using a 0.18-${\mu}m$ CMOS cell library with 100 MHz clock has 284,409 gates and RAM of 62,976 bits, and it is verified by FPGA implementation. The estimated performance depending on code rate and block length is about 82~218 Mbps at 100 MHz@1.8V.

Synthesis and Secretion of Mutant Mannose-Binding Lectin (돌연변이 Mannose-binding Lectin 합성과 세포 병리적 연구)

  • Jang, Ho-Jung;Chung, Kyung Tae
    • Journal of Life Science
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    • v.23 no.3
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    • pp.347-354
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    • 2013
  • Innate immunity is the ability to differentiate infectious agents from self. The innate immune system is comprised of a complicated network of recognition and effector molecules that act together to protect the host in the early stage of an infectious challenge. Mannose-binding lectin (MBL or mannose-binding protein, MBP) belongs to the family of $Ca^{2+}$-dependent lectins (C-type lectin with a collagen-like domain), which are considered an important component of innate immunity. While it is associated with increased risk and severity of infections and autoimmunity, the most frequent immuno-deficiency syndrome was reported to be low MBL level in blood. Deficiency of human MBL is caused by mutations in the coding region of the MBL gene. Rat homologue gene of human MBL gene was used to study functions of wild type and mutant MBL proteins. Although extensive studies have yielded the structural information of MBL, the functions of MBL, especially mutant MBL, still require investigation. We previously reported the cloning of rat wild-type MBL gene and the production of a truncated form of MBL protein and its antibody. Here, we present the cloning of mutant MBL cDNA in collagen-like domain (R40C, G42D, and G45E) using site-directed mutagenesis and differential behaviors of wild type and mutant MBL in cells. The major difference between wild type and mutant MBL was that while wild type MBL was secreted, mutant MBL was inhibited for secretion, retained in endoplasmic reticulum, and still functioned as a lectin.

Isolation and Characterization of a Novel Calcium/Calmodulin-Dependent Protein Kinase, AtCK, from Arabidopsis

  • Jeong, Jae Cheol;Shin, Dongjin;Lee, Jiyoung;Kang, Chang Ho;Baek, Dongwon;Cho, Moo Je;Kim, Min Chul;Yun, Dae-Jin
    • Molecules and Cells
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    • v.24 no.2
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    • pp.276-282
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    • 2007
  • Protein phosphorylation is one of the major mechanisms by which eukaryotic cells transduce extracellular signals into intracellular responses. Calcium/calmodulin ($Ca^{2+}/CaM$)-dependent protein phosphorylation has been implicated in various cellular processes, yet little is known about $Ca^{2+}/CaM$-dependent protein kinases (CaMKs) in plants. From an Arabidopsis expression library screen using a horseradish peroxidase-conjugated soybean calmodulin isoform (SCaM-1) as a probe, we isolated a full-length cDNA clone that encodes AtCK (Arabidopsis thaliana calcium/calmodulin-dependent protein kinase). The predicted structure of AtCK contains a serine/threonine protein kinase catalytic domain followed by a putative calmodulin-binding domain and a putative $Ca^{2+}$-binding domain. Recombinant AtCK was expressed in E. coli and bound to calmodulin in a $Ca^{2+}$-dependent manner. The ability of CaM to bind to AtCK was confirmed by gel mobility shift and competition assays. AtCK exhibited its highest levels of autophosphorylation in the presence of 3 mM $Mn^{2+}$. The phosphorylation of myelin basic protein (MBP) by AtCK was enhanced when AtCK was under the control of calcium-bound CaM, as previously observed for other $Ca^{2+}/CaM$-dependent protein kinases. In contrast to maize and tobacco CCaMKs (calcium and $Ca^{2+}/CaM$-dependent protein kinase), increasing the concentration of calmodulin to more than $3{\mu}M$ suppressed the phosphorylation activity of AtCK. Taken together our results indicate that AtCK is a novel Arabidopsis $Ca^{2+}/CaM$-dependent protein kinase which is presumably involved in CaM-mediated signaling.

Soluble Prokaryotic Expression and Purification of Bioactive Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand

  • Do, Bich Hang;Nguyen, Minh Tan;Song, Jung-A;Park, Sangsu;Yoo, Jiwon;Jang, Jaepyeong;Lee, Sunju;So, Seoungjun;Yoon, Yejin;Kim, Inki;Lee, Kyungjin;Jang, Yeon Jin;Choe, Han
    • Journal of Microbiology and Biotechnology
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    • v.27 no.12
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    • pp.2156-2164
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    • 2017
  • Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered as an antitumor agent owing to its ability to induce apoptosis of cancer cells without imparting toxicity toward most normal cells. TRAIL is produced in poor yield because of its insoluble expression in the cytoplasm of E. coli. In this study, we achieved soluble expression of TRAIL by fusing maltose-binding protein (MBP), b'a' domain of protein disulfide isomerase (PDIb'a'), or protein disulfide isomerase at the N-terminus of TRAIL. The TRAIL was purified using subsequent immobilized metal affinity chromatography and amylose-binding chromatography, with the tag removal using tobacco etch virus protease. Approximately 4.5 mg of pure TRAIL was produced from 125 ml flask culture with a purification yield of 71.6%. The endotoxin level of the final product was $0.4EU/{\mu}g$, as measured by the Limulus amebocyte lysate endotoxin assay. The purified TRAIL was validated and shown to cause apoptosis of HeLa cells with an $EC_{50}$ and Hill coefficient of $0.6{{\pm}}0.03nM$ and $2.41{\pm}0.15$, respectively. The high level of apoptosis in HeLa cells following administration of purified TRAIL indicates the significance and novelty of this method for producing high-grade and high-yield TRAIL.

A Study on integrated to communication and broadcasting cable telecommunication Structure for Digital Conversion (통방통합 유선전송망의 디지털 전환을 위한 전송망근조에 관한 연구)

  • Sung yong-seok;Jin Yong-Ok
    • Proceedings of the Korean Society of Broadcast Engineers Conference
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    • 2004.11a
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    • pp.31-35
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    • 2004
  • 정보통신 기술의 발달과 디지털 방송의 시작, 뉴미디어의 출현으로 인한 통신과 방송의 융합은 가속화되고 있다. 또한 2010년까지 광대역통신망 BcN과 홈네트워크 구축을 위한 정부정책이 실행 중에 있다. 100Mbps의 전송속도를 구현해야 하는 광대역 통신망(BcN)을 위해 기존 인터넷 백본 1)망은 잘 구축이 되어 있으나 가입자까지의 망 구조에 많은 문제점을 앉고 있다. 기존 전화국을 이용한 XDSL과 지역 SO를 활용한 Cable Modem의 경우 병목현상과 이론상 속도 또한 BcN과 통방통합이 요구하는 50-100Mhz의 전송속도를 만족하지 못한다. 새로운 망 구조를 구축하기 위해 많은 비용과 시간의 소요가 예상된다 가입자 망 구축에 따른 많은 방법과 이론이 제시되고 있다. 똔 논문에선 지역 SO를 활용하여 가입자까지 망을 통방통합과 BcN에 적합한 가입자 망을 새롭게 구성하는 것을 목표로 한다. 먼저 지역 50의 망을 활용하기 위해선 기존 KT와 파워콤의 COF(Glass Optical Fiber)망과 지역 케이블 SO의 HFC 망을 이용하기에는 동축케이블 망의 물리적 특성에 따른 한계로 통방통합과 BcN에 부적합하다. Tree And Branch 구조의 HFC망 대신 $SMF^{2)}$의 기존 SO의 자가망을 새롭게 설계하고 광분배망 기술인 $E-PON^{3)}$방식을 접목시켜 최대한 동축망을 사용하지 않고 굴곡 특성에 약한 $FOG^{4)}$의 특성을 극복하기 위해 $POF^{5)}$망을 이용하여 댁내 홈게이트웨이까지 연결하는 방식으로 지역 SO를 거점으로 활용하여 댁내까지 FHHT와 홈 네트워크까지의 가입자 망을 새롭게 구성하고자 한다. 저장의 효율성을 위해 이진 포멧인 IPMP화된 MP4 파일을 생성할 수 있다.으로써, 에러 이미지가 가지고 있는 엔트로피에 좀 근접하게 코딩을 할 수 있게 되었다. 이 방법은 실제로 Arithmetic Coder를 이용하는 다른 압축 방법에 그리고 적용할 수 있다. 실험 결과 압축효율은 JPEG-LS보다 약 $5\%$의 압축 성능 개선이 있었으며, CALIC과는 대등한 압축률을 보이며, 부호화/복호화 속도는 CALIC보다 우수한 것으로 나타났다.우 $23.87\%$($18.00\~30.91\%$), 갑폭 $23.99\%$($17.82\~30.48\%$), 체중 $91.51\%$($58.86\~129.14\%$)이였으며 성장율은 사육 온도구간별 차는 없었다.20 km 까지의 지점들(지점 2에서 지점 6)에서 매우 높은 값을 보이며 이는 조석작용으로 해수와 담수가 강제혼합되면서 표층퇴적물이 재부유하기 때문이라고 판단된다. 영양염류는 월별로 다소의 차이는 있으나, 대체적으로 지점 1과 2에서 가장 낮고, 상류로 갈수록 점차 증가하며 지점 7 상류역이 하류역에 비해 높은 농도이다. 월별로는 7월에 규산염, 용존무기태질소 및 암모니아의 농도가 가장 높은 반면에 용존산소포화도는 가장 낮다. 그러나 지점 14 상류역에서는 5월에 측정한 용존무기태질소, 암모니아, 인산염 및 COD 값이 7월보다 다소 높거나 비슷하다. 한편 영양염류와 COD값은 대체적으로 8월에 가장 낮으나 용존산소포화도는 가장 높다.출조건은 $100^{\circ}C$에서 1분간의 고온단시간 추출이 적합하였다. 증가를 나타내었는데

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A Study on Nonlinear Distortion Analysis of Power Amplifier using the OFDM for WLAN System (무선랜 시스템에서 OFDM 방식을 사용한 전력증폭기의 비선형 왜곡분석에 관한 연구)

  • Oh Chung-Gyun;Kim Dong-Ok
    • Journal of The Institute of Information and Telecommunication Facilities Engineering
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    • v.2 no.4
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    • pp.42-51
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    • 2003
  • In this paper, we are going to analyze on relation of an output spectrum along phase distortion of power amplifier in wireless LAN system, and then considered an ACPR characteristic of power amplifier and consideration of an OFDM method for this. Also, we did implementation for OFDM modulation and transmission section of an IEEE 802.11a standard to have transmission speed of the maximum 54Mbps in order to know an OFDM modulation method and relation of non-linear characteristic of power amplifier. The non-linear characteristic of power amplifier did modeling with AM-to-AM and AM-to-PM, and we analyzed an output spectrum characteristic along phase distortion composed input signal supply for power amplifier. When output spectrum analysis results phase distortion increased, and an AM-to-PM characteristic of power amplifier in 5 degrees, the output spectrum was satisfied with a demand spectrum in P1 dB, but 10-20 degrees were able to confirm what cannot be satisfied with a demand spectrum in phase distortion. Also, an output spectrum of power amplifier by frequency re-growth generated by a non-linear characteristic of power amplifier did not satisfied in P1dE. therefore, a back-off value was requested according to an AM-to-PM distortion degree, and smaller back-off value were able to know what demand became in case of modulation section that used OFDM.

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Isolation and Characterization of Lactic Acid Bacteria from Fermented Goat Milk in Tajikistan

  • Cho, Gyu-Sung;Cappello, Claudia;Schrader, Katrin;Fagbemigun, Olakunle;Oguntoyinbo, Folarin A.;Csovcsics, Claudia;Rosch, Niels;Kabisch, Jan;Neve, Horst;Bockelmann, Wilhelm;Briviba, Karlis;Modesto, Monica;Cilli, Elisabetta;Mattarelli, Paola;Franz, Charles M.A.P
    • Journal of Microbiology and Biotechnology
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    • v.28 no.11
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    • pp.1834-1845
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    • 2018
  • The lactobacilli associated with a fermented goat milk product from Tajikistan were isolated to characterize their technological properties and antibiotic resistances in order to assess their suitability for development as starter cultures. In this study, twenty three strains were identified by 16S rRNA sequencing as typical dairy-associated lactic acid bacterial strains, i.e. L. plantarum, L. pentosus, L. delbrueckii, L. helveticus and L. paracasei. These strains were generally susceptible to most antibiotics tested in this study and this allowed a selection of strains as safe starters. The draft genomes of four representative strains were sequenced and the number of contigs of the four assembled genomes ranged from 51 to 245 and the genome sizes ranged from 1.75 to 3.24 Mbp. These representative strains showed differences in their growth behavior and pH-reducing abilities in in vitro studies. The co-inoculation of these Lactobacillus spp. strains together with a yeast Kluyveromyces marxianus MBT-5698, or together with the yeast and an additional Streptococcus thermophilus MBT-2, led to a pH reduction to 3.4 after 48 h. Only in the case of fermentation inoculated with the co-culture, the viscosity of the milk increased noticeably. In contrast, fermentations with single strains did not lead to gelation of the milk or to a decrease in the pH after 24h. The results of this study provide a comprehensive understanding of the predominant lactobacilli related to Tajikistani fermented milk products.

Study on the soluble exoression of recombinant human eoidermal growth factor using various fusion oartners in Escherichia coli (재조합 대장균에서 다양한 융합 파트너를 이용한 인간 상피세포성장인자의 발현 연구)

  • Kim, Byung-Lip;Baek, Jung-Eun;Kim, Chun-Sug;Lee, Hyeok-Weon;Ahn, Jung-Oh;Lee, Hong-Weon;Jung, Joon-Ki;Lee, Eun-Gyo;Kim, In-Ho
    • KSBB Journal
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    • v.23 no.3
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    • pp.205-212
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    • 2008
  • The efficient soluble expression of human epidermal growth factor (hEGF) was achieved by using functional fusion partners in cytoplasm and periplasm of Escherichia coli (E. coli). hEGF was over-expressed in inactive inclusion body form in cytoplasm of E. coli due to improper disulfide bond formation and hydrophobic interaction, yielding about 5.9 mg/L in flask culture. Six functional fusion partners were introduced by linking to N-terminal part of hEGF gene for the high-level expression of soluble and active hEGF in cytoplasm and peri plasm region. Three fusion partners for cytoplasmic expression such as acidic tail of synuclein (ATS), thioredoxin (Trx) and lipase, and three fusion partners for periplasmic expression such as periplasmic cystein oxidoreductases (DsbA and DsbC) and maltose binding protein (MBP) were investigated. hEGF fused with ATS and DsbA showed over 90% of solubility in cytoplasm and periplasm, respectively. Especially DsbA was found to be an efficient fusion partner for soluble and high-level expression of hEGF, yielding about 18.1 mg/L and three-fold higher level compared to that of insoluble non-fusion hEGF in cytoplasm. Thus, heterologous proteins containing complex disulfide bond and many hydrophobic amino acids can effectively be produced as an active form in E. coli by introducing a suitable peptide or protein.