• Title/Summary/Keyword: cell identity

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Biochemical and Molecular Analysis of OsPAP1: A Phosphate Starvation Induced Purple Acid Phosphatase Gene from Rice

  • Hur, Yeon Jae;Yi, Young Byung;Kim, Tae Ho;Kim, Doh Hoon
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.455-462
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    • 2010
  • Purple acid phosphatase is important for phosphorus remobilization in plants, but its role in plant adaptation to low phosphorus availability is not known. The cDNA encoding O. sativa purple acid phosphatase (OsPAP1) has 1008 bp with an open reading frame of 335 amino acid residues. The amino acid sequence of OsPAP1 cDNA shows of 50-51% identity with other plant purple acid phosphatases. OsPAP1 was expressed in rice plants and in cell cultures in the absence of phosphate ($P_i$). The expression was organ-specific with the strongest expression in $P_i$-deprived roots. Functional expression of the OsPAP1 gene in the transgenic Arabidopsis line was confirmed by northern and western blot analysis. OsPAP1 overexpression lines had higher phosphatase activity than wild-type. Overexpression of OsPAP1 in Arabidopsis plants resulted in increased Pi accumulation under Pi sufficient condition. These results show that the OsPAP1 gene represents more efficient $P_i$ uptake and can be used to develop new transgenic dicotyledonous plants.

Efficient Proxy Re-encryption Scheme for E-Voting System

  • Li, Wenchao;Xiong, Hu
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.15 no.5
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    • pp.1847-1870
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    • 2021
  • With the development of information and communication technologies, especially wireless networks and cell phones, the e-voting system becomes popular as its cost-effectiveness, swiftness, scalability, and ecological sustainability. However, the current e-voting schemes are faced with the problem of privacy leakage and further cause worse vote-buying and voter-coercion problems. Moreover, in large-scale voting, some previous e-voting system encryption scheme with pairing operation also brings huge overhead pressure to the voting system. Thus, it is a vital problem to design a protocol that can protect voter privacy and simultaneously has high efficiency to guarantee the effective implementation of e-voting. To address these problems, our paper proposes an efficient unidirectional proxy re-encryption scheme that provides the re-encryption of vote content and the verification of users' identity. This function can be exactly applied in the e-voting system to protect the content of vote and preserve the privacy of the voter. Our proposal is proven to be CCA secure and collusion resistant. The detailed analysis also shows that our scheme achieves higher efficiency in computation cost and ciphertext size than the schemes in related fields.

A Study on the Preparation of Standardized Operation Criteria for Enhancement of Safety and Convenience of Mobile Electronic Notice Service

  • JongBae, Kim
    • International Journal of Advanced Culture Technology
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    • v.10 no.4
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    • pp.547-554
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    • 2022
  • Due to the expansion of non-face-to-face services, the demand for user identification for mobile devices is increasing. Recently, mobile resident registration cards, mobile driver's licenses, etc. are installed in mobile phones and used for user identification and authentication services. In order to identify a user online, unique identification information of the online user is required. In particular, in order to provide information only to online users, it is necessary to accurately deliver information to a mobile device owned by the user. To make this service possible, it was realized with the advent of mobile electronic notice service. However, the identification of online service users and information on mobile devices owned or subscribed by the relevant users require safe management as personal information, and it is also necessary to increase the convenience of online service users. In this paper, we propose an operating standard for providing a mobile electronic notice service that sends electronic notice using a mobile device owned by the user. The mobile electronic notice service is a service that provides notices expressed in electronic information to the recipient's cell phone, mobile app, e-mail, etc. Therefore, as the use of mobile electronic notification service increases and the provision and use of connecting information to identify users increases, it is necessary to expand the mobile electronic notification service while safely protecting users' personal information.

Increased osteopontin expression in activated glial cells in experimental autoimmune encephalomyelitis (자가면역성 뇌척수염 흰쥐의 활성화된 신경아교세포에서 증가된 osteopontin의 발현)

  • Park, Suk-jae;Hwang, In-sun;Kim, Gyu-beom;Shin, Tae-kyun;Jee, Young-heun
    • Korean Journal of Veterinary Research
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    • v.46 no.3
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    • pp.177-184
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    • 2006
  • Experimental autoimmune encephalomyelitis (EAE) is a disease model of multiple sclerosis (MS) that is characterized by remittance and relapse of the disease and autoimmune and demyelinating lesions in the central nervous system (CNS). Autoimmune inflammation is maintained by secretion of a large number of protein. Previous studies have suggested that transcripts encoding osteopontin (OPN) are frequently detected in the mRNA population of MS plaques. To elucidate the functional role of OPN in initiation and development of EAE, we examined the expression and localization of OPN in the spinal cord during acute EAE. We demonstrated that OPN significantly increased at the early stage of EAE and slightly declined thereafter by western blot analysis. An immunohistochemical study revealed that OPN was constitutively expressed in some glial cells (microglia, astrocytes) of white matter and neurons in the CNS of control rats. OPN expression was shown to be increased in the same cells at the early and peak stage of EAE. To identity cells expressing OPN by double-immunofluorescence labeling, we labeled rat spinal cord sections for OPN with a monoclonal OPN antibody and with mAbs for astrocyte (GFAP), microglia/macrophage (OX42)-specific markers. The major cell types of OPN-expressing cells were activated astrocytes and microglia in the adjacent inflammatory lesions. Interestingly, OPN was mainly expressed in the end feet of astrocytes around vascular cell adhesion molecule-1 (VCAM-1) expressing endothelial cells of CNS blood vessel. These findings suggest that increased levels of OPN in activated glial cell may play an important role in the recruitment of inflammatory cells into the CNS parenchyma during EAE.

Cloning and functional expression of a cecropin-A gene from the Japanese oak silkworm, Antheraea yamamai (천잠 cecropin-A 유전자 클로닝 및 재조합 발현)

  • Kim, Seong-Ryul;Choi, Kwang-Ho;Kim, Sung-Wan;Goo, Tae-Won;Hwang, Jae-Sam
    • Journal of Sericultural and Entomological Science
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    • v.52 no.1
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    • pp.45-51
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    • 2014
  • A cecropin-A gene was isolated from the immunized larvae of the Japanese oak silkworm, Antheraea yamamai and designed Ay-CecA. The complete Ay-CecA cDNA consists of 419 nucleotides with 195 bp open reading frame encoding a 64 amino acid precursor that contains a putative 22-residue signal peptide, a 4-residue propetide and a 37-residue mature peptide with a theoretical mass of 4046.81. The deduced amino acid sequence of the peptide evidenced a significant degree of identity (62 ~ 78% identity) with other lepidopteran cecropins. Like many insect cecropin, Ay-CecA also harbored a glycine residue for C-terminal amidation at the C-end, which suggests potential amidation. To understand this peptide better, we successfully expressed bioactive recombinant Ay-CecA in Escherichia coli that are highly sensitive to the mature peptide. For this, we fused mature Ay-CecA gene with insoluble protein ketosteroid isomerase (KSI) gene to avoid the cell death during induction. The fusion KSI-CecA protein was expressed as inclusion body. The expressed fusion protein was purified by Ni-NTA immobilized metal affinity chromatography (IMAC), and cleaved by cyanogen bromide (CNBr) to release recombinant Ay-CecA. The purified recombinant Ay-CecA showed considerably antibacterial activity against Gram-negative bacteria, E. cori ML 35, Klebsiella pneumonia and Pseudomonas aeruginosa. Our results proved that this peptide with a potent antibacterial activity may play a role in the immune response of Japanese oak silkworm.

A Parentage Test using Indel, Microsatellite Markers and Genotypes of MC1R in the Jeju Black Cattle Population (제주 흑우 집단에서 Indel, Microsatellite 마커와 MC1R 유전자형을 이용한 친자 확인)

  • Han, Sang Hyun;Cho, Sang-Rae;Cho, In-Cheol;Cho, Won-Mo;Kim, Sang-Geum;Yang, Sung-Nyun;Kang, Yong-Jun;Park, Yong-Sang;Kim, Young-Hoon;Park, Se-Phil;Kim, Eun-Young;Lee, Sung-Soo;Ko, Moon-Suck
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.207-213
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    • 2013
  • This study was carried out to examine a molecular marker system for parentage test in Jeju Black cattle (JBC). Based on the preliminarily studies, we finally selected for construction of a novel genetic marker system for molecular traceability, identity test, breed certification, and parentage test in JBC and its related industrial populations. The genetic marker system had eight MS markers, five indel markers, and two single nucleotide polymorphisms (SNPs; g.G299T and g.del310G) within MC1R gene which is critical to verify the breed specific genotypes for coat color of JBC differing from those of exotic black cattle breeds such as Holstein and Angus. The results showed lower level of a combined non-exclusion probability for second parent (NE-P2) of $4.1202{\times}10^{-4}$ than those previously recommended by International Society of Animal Genetics (ISAG) of $5.000{\times}10^{-4}$ for parentage, and a combined non-exclusion probability for sib identity (NE-SI) of $2.679{\times}10^{-5}$. Parentage analysis has been successfully identified the JBC offspring in the indigenous population and cattle farms used the certified AI semens for production using the JBC-derived offspring for commercial beef. This combined molecular marker system will be helpful to supply genetic information for parentage test and traceability and to develop the molecular breeding system for improvement of animal productivity in JBC population.

Cloning of SNAS-25 Gene from Rat Brain cDNA Library (Rat Brain cDNA Library로부터 SNAP-25 유전자의 클로닝)

  • Cho, Ae-Ri;Ji, Young-Mi;Yoo, Min;Lee, Soon-Chul;Yoo, Kwan-Hee
    • Biomedical Science Letters
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    • v.6 no.1
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    • pp.11-17
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    • 2000
  • SNAP-25 was first investigated as a neuron-specific protein preferentially expressed in CA3 pyramidal neurons of mouse hippocampus. It is a presynaptic plasma membrane protein in the nerve cell and plays an important role in the synaptic vesicle membrane docking and fusion pathway. We have recently isolated SNAP-25 cDNA from a rat brain cDNA library using a probe of Z2 cDNA. It consisted of 2,101 bp and an open reading frame (ORF) was identified between nucleotides (nt) 209 and 827. The AUG codon (nt 209∼211) was surrounded by CTACCATGG, which corresponded to the consensus sequence of ribosomal binding site. The ORF was terminated by TAA (nt 827∼829) to encode a polypeptide of 206 amino acid residues. The 3'-untranslated region contained two extensive stretches of repeated (CA)28 and (CA)19 at positions 925∼980 and 1645∼1682. It is noteworthy that cysteine residues were clustered in the span of amino acid residues 84∼991 : Cys-Gly-Leu-Cys-Val-Cys-Pro-Cys. Rat SNAP-25 showed 88% and 97% identity in nucleotide sequences to that of human and mouse, respectively. Amino acid sequence of rat SNAP-25 showed 100% identity to that of mouse and human SNAP-21.

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Sequencing of cDNA Clones Expressed in Adipose Tissues of Korean Cattle

  • Bong, J.J.;Tong, K.;Cho, K.K.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.4
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    • pp.483-489
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    • 2005
  • To understand the molecular mechanisms that regulate intramuscular fat deposition and its release, cDNA clones expressed in adipose tissues of Korean cattle were identified by differential screening from adipose tissue cDNA library. By partial nucleotide sequencing of 486 clones and a search for sequence similarity in NCBI nucleotide databases, 245 clones revealed unique clones. By a functional grouping of the clones, 14% of the clones were categorized to metabolism and enzyme-related group (stearoyl CoA desaturase, lactate dehydrogenase, fatty acid synthase, ATP citrate lyase, lipoprotein lipase, acetyl CoA synthetase, etc), and 6% to signal transduction/cell cycle-related group (C/EBP, cAMP-regulated phosphoprotein, calmodulin, cyclin G1, cyclin H, etc), and 4% to cytoskeleton and extracellular matrix components (vimentin, ankyrin 2, gelosin, syntenin, talin, prefoldin 5). The obtained 245 clones will be useful to study lipid metabolism and signal transduction pathway in adipose tissues and to study obesity in human. Some clones were subjected to full-sequencing containing open reading frame. The cDNA clone of bovine homolog of human prefoldin 5 gene had a total length of 959 nucleotides coding for 139 amino acids. Comparison of the deduced amino acid sequences of bovine prefoldin 5 with those of human and mouse showed over 95% identity. The cDNA clone of bovine homolog of human ubiquitin-like/S30 ribosomal fusion protein gene had a total length of 484 nucleotides coding for 133 amino acids. Comparison of the deduced amino acid sequences of bovine ubiquitin-like/S30 ribosomal fusion protein gene with those of human, rat and mouse showed over 97% identity. The cDNA clone of bovine homolog of human proteolipid protein 2 mRNA had a total length of 928 nucleotides coding for 152 amino acids. Comparison of the deduced amino acid sequences of bovine proteolipid protein 2 with those of human and mouse showed 87.5% similarity. The cDNA clone of bovine homolog of rat thymosin beta 4 had a total length of 602 nucleotides coding for 44 amino acids. Comparison of the deduced amino acid sequences of bovine thymosin beta 4 gene with those of human, mouse and rat showed 93.1% similarity. The cDNA clone of bovine homolog of human myotrophin mRNA had a total length of 790 nucleotides coding for 118 amino acids. Comparison of the deduced amino acid sequences of bovine myotrophin gene with those of human, mouse and rat showed 83.9% similarity. The functional role of these clones in adipose tissues needs to be established.

Cloning a Mannanase 26AT Gene from Paenibacillus woosongensis and Characterization of the Gene Product (Paenibacillus woosongensis으로부터 Mannanase 26AT 유전자의 클로닝과 유전자 산물의 분석)

  • Yoon, Ki-Hong
    • Journal of Life Science
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    • v.27 no.9
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    • pp.1003-1010
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    • 2017
  • An open reading frame coding for mannanase predicted from the partial genomic sequence of Paenibacillus woosongensis was cloned into Escherichia coli by polymerase chain reaction amplification, and completely sequenced. This mannanase gene, designated man26AT, consisted of 3,162 nucleotides encoding a polypeptide of 1,053 amino acid residues. Based on the deduced amino acid sequence, Man26AT was identified as a modular enzyme, which included a catalytic domain belonging to the glycosyl hydrolase family 26 and two carbohydrate-binding modules, CBM27 and CBM11. The amino acid sequence of Man26AT was homologous to that of several putative mannanases, with identity of 81% for P. ihumii and identity of less than 57% for other strains of Paenibacillus. A cell-free extract of recombinant E. coli carrying the man26AT gene showed maximal mannanase activity at $55^{\circ}C$ and pH 5.5. The enzyme retained above 80% of maximal activity after preincubation for 1 h at $50^{\circ}C$. Man26AT was comparably active on locust bean gum (LBG), galactomanan, and kojac glucomannan, whereas it did not exhibit activity on carboxymethylcellulose, xylan, or para-nitrophenyl-${\beta}$-mannopyranoside. The common end products liberated from mannooligosaccharides, including mannotriose, mannotetraose, mannopentaose, and mannohexaose, or LBG by Man26AT were mannose, mannobiose, and mannotriose. Mannooligosacchrides larger than mannotriose were found in enzymatic hydrolyzates of LBG and guar gum, respectively. However, Man26AT was unable to hydrolyze mannobiose. Man26AT was intracellularly degraded into at least three active proteins with different molecular masses by zymogram.

Genetic Stock Identification of Spotted Flounder, Verasper variegatus from Yeocheun, Korea (범가자미에 대한 유전학적 동정)

  • KIM Kyung Kil;KIM Yoon;NAM Yoon Kwan;KIM Dong Soo
    • Journal of Aquaculture
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    • v.6 no.3
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    • pp.221-233
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    • 1993
  • Cell size, DNA content, chromosome and PCR-based mitochondrial 12S rRNA gene analyses were conducted to obtain basic informations for genetic stock identification of spotted flounder (Verasper variegatus) from Yeocheun, Korea. The mean erythrocytic and nuclear volumes of spotted flounder were $211.10{\mu}m^3$ and $23.03{\mu}m^3$, respectively. The haploid DNA content of this species was 0.79 pg/cell which correspond to $46.5\%$ of carp and to $22.6\%$ of mammals. Spotted flounder had the 2n = 46 acrocentric chromosomes but no heteromorphic sex chromosomes was found. Mitochondrial DNA gene for 12S ribosomal RNA was amplified by polymerase chain reaction (PCR) and the PCR products were subjected to digestion with 15 restriction endonucleases. Restriction enzyme analyses revealed that Ava I, Mae II, Sma I and Xba I had one restriction site in the mitochondrial 12S rRNA gene segment of spotted flounder, while Mae I had two. Segments of 12S rRNA gene from mitochondria in spotted flounder were sequenced and compared with channel catfish and human as controls. The 12S rRNA gene of this species was more similar to that of channel catfish than to human's.

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