• Title/Summary/Keyword: Functional characterization

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Functional characterization of a CCCH type zinc-finger protein gene OsZF2 by ectopic overexpression of the gene in rice (과발현 형질전환벼에서 CCCH type zinc-finger protein 유전자 OsZF2 기능 분석)

  • Lee, Jung-Sook;Yoon, In-Sun;Yoon, Ung-Han;Lee, Gang-Seob;Byun, Myung-Ok;Suh, Seok-Chul
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.23-29
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    • 2009
  • We have previously isolated a CCCH type zinc-finger protein gene, OsZF2 (Oryza sativa Zinc Finger 2), from the cold-treated rice cDNA library. To investigate the potential role of OsZF2, transgenic rice lines over-expressing OsZF2 under the control of CaMV 35S promoter have been developed through Agrobacterium-mediated transformation. Elevated level of OsZF2 transcripts was confirmed by RNA gel blot analysis in transgenic rice. Under the 100 mM NaCl condition, the transgenic rice showed significantly enhanced growth rate in terms of shoot length and fresh weight, implicating that OsZF2 is likely to be involved in salt response of rice. In the field condition, however, the transgenic rice showed a dwarf phenotype and flowering time was delayed. Genome expression profiling analysis of transgenic plants using the 20K NSF rice oligonucleotide array revealed many up-regulated genes related to stress responses and signaling pathways such as chaperone protein dnaJ 72, salt stress-induced protein, PR protein, disease resistance proteins RPM1 and Cf2/Cf5 disease resistance protein, carbohydrate/ sugar transporter, OsWAK kinase, brassinosteroid LRR receptor kinase, and jasmonate O-methyltransferase. These data suggest that the CCCH type zinc-finger protein OsZF2 is a upstream transcriptional factor regulating growth and stress responsiveness of rice.

Characterization of Bovine Brucellosis in Korean Native Cattle by Means of Immunohistochemistry and Proteomics (면역조직 화학법 및 단백질체 변화 분석을 통한 한우에서 발생한 브루셀라증의 특성)

  • Jang, Seong-Jun;Do, Sun-Hee;Ki, Mi-Ran;Hong, Il-Hwa;Park, Jin-Kyu;Cho, Yu-Jeong;Park, Sang-Joon;Kim, Tae-Hwan;Kwak, Dong-Mi;Jeong, Kyu-Shik
    • Journal of Life Science
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    • v.20 no.2
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    • pp.153-160
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    • 2010
  • This study was conducted to examine the utilization of immunohistochemistry using the bovine anti-brucella immunoglobulin G (IgG) antibody in the diagnosis of brucellosis and to develop a functional biomarker relation for the progress of the disease. Anti-brucella IgG antibody was purified from the affected bovine serum using an affinity chromatography. We performed our investigation on 17 cases of brucellosis and 19 control cases with negative Rose-Bengal test results. Our purified anti-brucella IgG antibody showed a positive immunoreactivity in cytoplasmic hepatocytes of the centrilobular region, and glomeruli and tubular epithelium of the kidney. The protein pattern of the affected liver versus control was analyzed by two-dimensional electrophoresis, showing a different expression pattern of proteins between the two. Five protein spots were up-regulated and another were five down-regulated in the brucellosis liver. Significant upregulaton of catalase and 3-hydroxyacyl-CoA dehydrogenase might be due to a compensatory reaction in response to the endotoxic shock of brucella. In conclusion, the anti-brucella IgG antibody may be a good tool for discriminative diagnosis of the affected tissues and proteomics data suggest new target proteins underlying a possible pathogenic mechanism of brucellosis.

Molecular cloning and characterization of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (CaHDR) from Camptotheca acuminata and its functional identification in Escherichia coli

  • Wang, Qian;Pi, Yan;Hou, Rong;Jiang, Keji;Huang, Zhuoshi;Hsieh, Ming-shiun;Sun, Xiaofen;Tang, Kexuan
    • BMB Reports
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    • v.41 no.2
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    • pp.112-118
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    • 2008
  • Camptothecin is an anti-cancer monoterpene indole alkaloid. The gene encoding 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (designated as CaHDR), the last catalytic enzyme of the MEP pathway for terpenoid biosynthesis, was isolated from camptothecin-producing Camptotheca acuminata. The full-length cDNA of CaHDR was 1686 bp encoding 459 amino acids. Comparison of the cDNA and genomic DNA of CaHDR revealed that there was no intron in genomic CaHDR. Southern blot analysis indicated that CaHDR belonged to a low-copy gene family. RT-PCR analysis revealed that CaHDR expressed constitutively in all tested plant organs with the highest expression level in flowers, and the expression of CaHDR could be induced by 100 ${\mu}M$ methyl-jasmonate (MeJA), but not by 100 mg/L salicylic acid (SA) in the callus of C. acuminata. The complementation of CaHDR in Escherichia coli ispH mutant MG1655 demonstrated its function.

Preparation and Characterization of Conducting Polymer Nanocomposites Including Graphene Oxide via In-situ Chemical Polymerization (제자리 화학중합을 통한 그래핀 옥사이드를 포함하는 전도성 고분자 나노복합체의 제조와 특성 분석)

  • Jeong, Yeonjun;Moon, Byung-Chul;Jang, Min-Chae;Kim, Yangsoo
    • Polymer(Korea)
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    • v.38 no.2
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    • pp.180-187
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    • 2014
  • Nanocomposites including graphene oxide (GO) and conducting polymers (PPy, PANI and PEDOT) were prepared via an in-situ chemical polymerization process, and their characteristic properties depending upon the change of conducting polymer (CP) content were analyzed. A confirmation was made on not only the functional groups formed in GO but also the presence of CP existent in the nanocomposites. The molecular interaction between GO and poly(4-styrene sulfonic acid) (PSSA) or CP in the nanocomposites was proposed. With the increase of PEDOT content in the GOPSS/PEDOT nanocomposite, the estimated value of $I_D/I_G$ regarding the Raman analysis of them was decreased and a major change of their Raman spectra characteristic peaks was observed. In the GO-PSS/PEDOT nanocomposite, PEDOT molecules made an exfoliation of GO-PSSA layers and thus they were intercalated among layers. Such a unique molecular morphology induced the highest electrical conductivity for the GO-PSS/PEDOT nanocomposite among three kinds of nanocomposites prepared in this study. It is also noted that the uniform morphology confirmed in this study helped a thermal stability improvement in the nanocomposite due to the presence of GO or GO-PSSA acting as a thermal barrier.

Molecular Characterization of a Korean Isolate of Human Norovirus, the Hu/NLV/Gunpo/2006/KO Strain (인체 노로바이러스의 한국분리주 Hu/NLV/Gunpo/2006/KO의 분자생물학적 특성)

  • Jeong, Ah-Yong;Yun, Sang-Im;Jee, Young-Mee;Kang, Yoon-Sung;Lee, Young-Min
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.105-111
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    • 2009
  • Norovirus (NV) with a variety of genotypes, a member of the family Caliciviridae, causes acute nonbacterial gastroenteritis in humans. We determined the nucleotide sequence of three open reading frames (ORFs) of a NV Korean strain and characterized the genetic relationship with others. The Korean strain designated Hu/NLV/Gunpo/2006/KO was isolated from the stool specimen of a 2-year-old female suffering from gastroenteritis. By performing reverse transcription and PCR amplification, three overlapping cDNAs were synthesized and used for direct sequencing. We found that like other NVs, this strain contains three ORFs: ORF1, 5,100 bp; ORF2, 1,647 bp; ORF3, 765 bp. Of 35 NVs, ORF1 had a level of genetic diversity lower than ORF2 and ORF3, of which the C-termini of the ORF2 and ORF3 showed a relatively high degree of genetic diversity. Phylogenetic analyses indicated that the Korean strain belonged to genogroup II, with Saitama U1, Gifu'96, Mc37, and Vietnam 026 being formed a single genetic cluster. The nucleotide sequence information of three ORFs of a NV Korean isolate will be useful not only for the development of a diagnostic tool and understanding of genetic relationship, but also provide important basic information for the functional analysis of their gene products.

Expression and Role of the System L Amino Acid Transporter in FOB Human Osteoblast Cells (사람 골모세포 FOB에서 아미노산 수송계 L의 발현 및 역할)

  • Kim, Chang-Hyun;Park, Joo-Cheol;Kim, Do Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.9
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    • pp.1367-1374
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    • 2005
  • Amino acid transporters play an important role in supplying nutrition to normal and cancer cells for cell proliferation. Amino acid transport system L is a major nutrient transport system responsible for the $Na^+$-independent transport of neutral amino acids including several essential amino acids. The system L is divided into two major subgroups, the L-tyre amino acid transporter 1 (LAT1) and the L-type amino acid transporter 2 (LAT2). In the present study, we have examined the expression and functional characterization of system L amino acid transporters in FOB human osteoblast cells. RT-PCR and western blot analysis have revealed that the FOB cells expressed LAT1, LAT2 together with their associating protein 4F2hc. The uptakes of $[^{14}C]_L$-leucine by FOB cells are $Na^+$-independent and almost completely inhibited by system L amino acid transporter selective inhibitor, 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH). These results suggest that the transport of neutral amino acids including several essential amino acids for cellular nutrition into the FOB human osteoblast cells is mediated by system L amino acid transporters.

Functional Characterization of the ${\alpha}$- and ${\beta}$-Subunits of a Group II Chaperonin from Aeropyrum pernix K1

  • Lee, Jin-Woo;Kim, Se Won;Kim, Jeong-Hwan;Jeon, Sung-Jong;Kwon, Hyun-Ju;Kim, Byung-Woo;Nam, Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.818-825
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    • 2013
  • We isolated and functionally characterized the ${\alpha}$- and ${\beta}$-subunits (ApCpnA and ApCpnB) of a chaperonin from Aeropyrum pernix K1. The constructed vectors pET3d-ApCpnA and pET21a-ApCpnB were transformed into E. coli Rosetta (DE3), BL21 (DE3), or CodonPlus (DE3) cells. The expression of ApCpnA (60.7 kDa) and ApCpnB (61.2 kDa) was confirmed by SDS-PAGE analysis. Recombinant ApCpnA and ApCpnB were purified by heat-shock treatment and anion-exchange chromatography. ApCpnA and ApCpnB were able to hydrolyze not only ATP, but also CTP, GTP, and UTP, albeit with different efficacies. Purified ApCpnA and ApCpnB showed the highest ATPase, CTPase, UTPase, and GTPase activities at $80^{\circ}C$. Furthermore, the addition of ApCpnA and ApCpnB effectively protected citrate synthase (CS) and alcohol dehydrogenase (ADH) from thermal aggregation and inactivation at $43^{\circ}C$ and $50^{\circ}C$, respectively. In particular, the addition of ATP or CTP to ApCpnA and ApCpnB resulted in the most effective prevention of thermal aggregation and inactivation of CS and ADH. The ATPase activity of the two chaperonin subunits was dependent on the salt concentration. Among the ions we examined, potassium ions were the most effective at enhancing the ATP hydrolysis activity of ApCpnA and ApCpnB.

Manufacturing of Mozzarella Cheese Analogues: A Review (MozzarellaCheeseAnalogue제조에 관한 연구: 총설)

  • Lee, Joon-Ha;Song, Kwang-Young;Seo, Kun-Ho;Yoon, Yoh-Chang
    • Journal of Dairy Science and Biotechnology
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    • v.30 no.2
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    • pp.111-117
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    • 2012
  • A number of recently developed cheese analogues that are available in dairy food markets are currently being produced for improving health or diet, and they include Mozzarella, Cheddar, American, Muenster, and other custom flavors. Cheese analogues have many benefits such as extended-and-improved shelf life, price stability, and functional qualities that include better texture, higher melting point, and better stretching properties. Various cheese analogues can now be made by using soybeans or soy protein products, gelatin, gum arabic, and other ingredients. Hence, in this study, on the basis of previously published studies, we recommend soy protein for cheese analogues, for improving the texture and flavor of cheese analogues. Moreover, the best conditions for making cheese analogues and the factors that affect the characterization of cheese analogues have been described in this paper.

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Hydrophobic Coating on Fish Feed Using Dielectric Barrier Discharge Plasma Polymerization (유전체장벽방전 플라즈마 중합을 이용한 양어 사료의 소수성 코팅)

  • Lee, Sang Baek;Hung, Trinhquang;Jo, Jin Oh;Jung, Jun Bum;Im, Tae Heon;Mok, Young Sun
    • Applied Chemistry for Engineering
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    • v.25 no.2
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    • pp.174-180
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    • 2014
  • A plasma hydrophobic coating on commercial fish feed was conducted to prolong the floating time of feed, thereby enhancing the feed consumption rate and reducing the contamination of water in fish farms. The hydrophobic coating on the fish feed was prepared using an atmospheric-pressure dielectric barrier discharge (DBD) plasma with hexamethyldisiloxane (HMDSO), toluene and n-hexane as the precursors. The effect of the parameters such as input power, precursor type and coating time on the coating performance were examined. The physicochemical properties of the coating layer were analyzed using a Fourier transform infrared (FTIR) spectrometer and a contact angle (CA) analyzer. The water CA increased after the coating preparation, indicating that the surface changed from hydrophilic to hydrophobic. The FTIR characterization revealed that the hydrophobic layer was comprised of functional groups such as $CH_3$, Si-O-Si and Si-C. As a result of the hydrophobic coating, the floating time of the fish feed increased from several seconds to 3 minutes, which suggested that the plasma coating method could be a viable means for practical applications. Compared to the water CA measured as soon as the coating layer was prepared, the 6-day aged sample exhibited a substantial CA increase, confirming the aging effect on the improvement of the hydrophobicity.

Functional characterization of Arabidopsis thaliana BLH 8, BEL1-Like Homeodomain 8 involved in environmental stresses (환경 스트레스에 관여하는 애기장대 BLH 8, BEL1-Like Homeodomain 8의 기능 분석)

  • Park, Hyeong-Cheol;Park, Ji-Young;Baek, Dong-Won;Yun, Dae-Jin
    • Journal of Plant Biotechnology
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    • v.38 no.2
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    • pp.162-168
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    • 2011
  • High salinity is a common stress condition that adversely affects plant growth and crop production. In response to various environmental stresses, plants activate a number of defense genes that function to increase the tolerance. To isolate Arabidopsis genes that are involved in abiotic stress responses, we carried out genetic screening using various mutant lines. Among them, the blh8 ($\b{B}$EL1-$\b{L}$ike $\b{H}$omeodomain $\underline{8}$) mutant specifically shows chlorotic phenotypes to ionic (specifically, $Na^+$ and $K^+$) stresses, but no differences in root growth. In addition, BLH8 is related to plant development and abiotic stress as predicted by a Graphical Gaussian Model (GGM) network program. It implies that BLH8 functions as a putative transcription factor related to abiotic stress responses. Collectively, our results show that gene network analysis is a useful tool for isolating genes involved in stress adaptation in plants.