• Title/Summary/Keyword: i-vectors

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Current Progress and Prospects of Reprogramming Factors - Stem Cells vs Germ Cells - (줄기세포와 생식세포에서 리프로그래밍 인자에 대한 최근 연구 동향과 전망)

  • Seo, You-Mi;Lee, Kyung-Ah
    • Development and Reproduction
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    • v.14 no.2
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    • pp.43-50
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    • 2010
  • Recently induced pluripotent stem (iPS) cells are derived from somatic cells by ectopic expression of several transcription factors (reprogramming factors) using technology of somatic cell reprogramming. iPS cells are able to selfrenew and differentiate into all type of cells in the body similarly to embryonic stem cells. Because iPS cells have advantages that can avoid immune rejection after transplantation and ethical issues unlike embryonic stem cells, research on iPS has made significant progress since the first report by Yamanaka in 2006. Nevertheless of many advantages of iPS, safer methods to introduce reprogramming factors into somatic cells must be developed due to safety concerns regarding viral vectors, and safer reprogramming factors to substitute the oncogenes should be evaluated for clinical application of iPS. Here we discuss the recent progress in reprogramming factors in embryonic stem cells, oocytes, and embryos, and discuss further research for finding new, more reliable and safer reprogramming factors.

Recombinant α and β Subunits of M.AquI Constitute an Active DNA Methyltransferase

  • Pinarbasi, Hatice;Pinarbasi, Ergun;Hornby, David
    • BMB Reports
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    • v.35 no.3
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    • pp.348-351
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    • 2002
  • AquI DNA methyltransferase, M.AquI, catalyses the transfer of a methyl group from S-adenosyl-L-methionine to the C5 position of the outermost deoxycytidine base in the DNA sequence 5'CYCGRG3'. M.AquI is encoded by two overlapping ORFs (termed $\alpha$ and $\beta$) instead of the single ORF that is customary for Class II methyltransferase genes. The structural organization of the M.AquI protein sequence is quite similar to that of other bacterial C5-DNA methyltransferases. Ten conserved motifs are also present in the correct order, but only on two polypeptides. We separately subcloned the genes that encode the $\alpha$ and $\beta$ subunits of M.AquI into expression vectors. The overexpressed His-fusion $\alpha$ and $\beta$ subunits of the enzyme were purified to homogeneity in a single step by Nickel-chelate affinity chromatography. The purified recombinant proteins were assayed for biological activity by an in vitro DNA tritium transfer assay. The $\alpha$ and $\beta$ subunits of M.AquI alone have no DNA methyltransferase activity, but when both subunits are included in the assay, an active enzyme that catalyses the transfer of the methyl group from S-adenosyl-L-methionine to DNA is reconstituted. We also showed that the $\beta$ subunit alone contains all of the information that is required to generate recognition of specific DNA duplexes in the absence of the $\alpha$ subunit.

Expression and Characterization of Human T-Cell Leukemia Virus Type-I Env and Gag Proteins

  • Son, Kyung-Hwa;Kim, Byong-Moon;Lee, Taik-You;Kim, Seong-Ryong;Kim, Kun-Soo;Lee, Jeong-Kug;Yang, Jai-Myung
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.311-317
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    • 1999
  • Human T-cell leukemia virus Type-I (HTLV-I) is etiologically associated with rare adult T-cell leukemia, a malignant T-cell disorder. cDNAs encoding p24 (gag), gp21(env), and pXII of HTLV-I were amplified by polymerase chain reaction (PCR) using the genomic DNA extracted from HUT102 cell line as a template. The amplified cDNAs were cloned into the Escherichia coli expression vectors and over-expression of the recombinant proteins were achieved by adding IPTG into the culture media in order to induce the promoter. The molecular weights of the recombinant p24, gp21, and pXII, determined by SDS-PAGE, were found to be approximately 28 kDa, 23 kDa, and 15 kDa, respectively. Reactivity of the recombinant proteins with human sera was tested by the immunoblot assay. The gp21 and p24 reacted against the sera obtained from HTLV-I-infected individuals but not against the sera obtained from normal persons. These results suggest that the recombinant proteins expressed in E. coli were recognized by antibodies in sera from HTLV-I infected patients. These recombinant proteins would be applicable for detecting the presence of antibodies against HTLV-I in human blood samples.

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Development and Verification of Digital EEG Signal Transmission Protocol (디지털 뇌파 전송 프로토콜 개발 및 검증)

  • Kim, Do-Hoon;Hwang, Kyu-Sung
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.38C no.7
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    • pp.623-629
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    • 2013
  • This paper presents the implementation result of the EEG(electroencephalogram) signal transmission protocol and its test platform. EEG measured by a dry-type electrode is directly converted into digital signal by ADC(analog-to-digital converter). Thereafter it is transferred DSP(digital signal processor) platform by $I^2C$(inter-integrated circuit) protocol. DSP conducts the pre-processing of EEG and extracts feature vectors of EEG. In this work, we implement the $I^2C$ protocol with 16 channels by using 10 or 12-bit ADC. In the implementation results, the overhead ratio for the 4 bytes data burst transmission measures 2.16 and the total data rates are 345.6 kbps and 414.72 kbps with 10-bit and 12-bit 1 ksps ADC, respectively. Therefore, in order to support a high speed mode of $I^2C$ for 400 kbps, it is required to use 16:1 and $(8:1){\times}2$ ratios for slave:master in 10-bit ADC and 12-bit ADC, respectively.

In Vivo Excision and Amplification of Large Human Genomic Segments Using Cre/loxP-and EBNA-1/oriP-mediated Machinery

  • Yoon, Young-Geol;Choi, Ja-Young;Kim, Jung-Min;Lee, Jun-Hyoung;Kim, Sun-Chang
    • BMB Reports
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    • v.34 no.4
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    • pp.322-328
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    • 2001
  • Excision and amplification of pre-determined, large genomic segments (taken directly from the genome of a natural host, which provides an alternative to conventional cloning in foreign vectors and hosts) was explored in human cells. In this approach, we devised a procedure for excising a large segment of human genomic DNA, the iNOS gene, by using the Cre/loxP system of bacteriophage P1 and amplifying the excised circles with the EBNA-1/oriP system of the Epstein-Barr virus. Two loxP sequences, each of which serves as a recognition site for recombinase Cre, were integrated unidirectionally into the 5'-UTR and 3'-UTR regions of the iNOS gene, together with an oriP sequence for conditional replication. The traps-acting genes cre and EBNA-1, which were under the control of a tetracycline responsive $P_{hcmv^*-1}$ promoter, were also inserted into the 5'-UTR and 3'-UTR regions of the iNOS gene, respectively, by homologous recombination. The strain carrying the inserted elements was stably maintained until the excision and amplification functions were triggered by the induction of cre and EBNA-1. Upon induction by doxycycline, Cre excised the iNOS gene that was flanked by two ZoxP sites and circularized it. The circularized iNOS gene was then amplified by the EBNA-1/oriP-system. With this procedure, approximately a 45.8-kb iNOS genomic fragment of human chromosome 17 was excised and successfully amplified in human cells. Our procedure can be used effectively for the sequencing of unclonable genes, the functional analysis of unknown genes, and gene therapy.

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Infections with Zoonotic Trematode Metacercariae in Yellowfin Goby, Acanthogobius flavimanus, from Coastal Areas of Republic of Korea

  • Sohn, Woon-Mok;Na, Byoung-Kuk;Cho, Shin-Hyeong;Ju, Jung-Won;Lee, Soon-Won;Seok, Won-Seok
    • Parasites, Hosts and Diseases
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    • v.56 no.3
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    • pp.259-265
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    • 2018
  • To investigate the infection status of zoonotic trematode metacercariae in yellowfin goby, Acanthogobius flavimanus, from coastal areas of the Republic of Korea (Korea), we examined total 344 gobies from 21 localities with an artificial digestion method from 2013 to 2017. The metacercariae of Stictodora lari were most frequently detected in 45.3% goby from 7 localities, i.e., Taean-gun (Chungcheongnam-do), Gochang-gun (Jeollabuk-do), Muan-gun, Shinan-gun, Haenam-gun (Jeollanam-do), Hadong-gun and Goseong-gun (Gyeongsangnam-do). Their infection rates were 90.0%, 66.7%, 46.7%, 8.0%, 3.3%, 26.7%, and 86.7% and intensities were 1,090, 6.2, 1.6 1.0, 2.0, 2.0, and 7.2 metacercariae per fish infected respectively. Heterophyopsis continua metacercariae were found in 38.2% goby from 6 localities, i.e., Gochang-gun, Muan-gun, Shinan-gun, Gangjin-gun, Boseong-gun (Jeollanam-do) and Goseong-gun, and their intensities were relatively low, 1-21 metacercariae. Stictodora fuscata metacercariae were detected in 61.3% goby from 4 localities, i.e., Taean-gun, Gochang-gun, Hadong-gun and Goseong-gun. Their infection rates were 90.0%, 53.3%, 5.9%, and 73.3% and intensities were 1,081, 3.1, 3.0, and 10.2 metacercariae per fish infected respectively. Heterophyes nocens metacercariae were found in 55.0% goby from Muan-gun and Shinan-gun. Total 3 metacercariae of Isthmiophora hortensis were detected in 2 (8.0%) gobies from Shinan-gun. Total 15 metacercariae of Centrocestus armatus were detected in 5 gobies (33.3%) from Gyeongpo-ho (ho means lake) in Gangneung-si, Gangwon-do. The present study suggests that yellowfin goby, A. flavimanus, acts as the infection sources of zoonotic intestinal flukes in western and southern coastal areas than in eastern coastal areas of Korea.

Homo- or Hetero-Dimerization of Muscarinic Receptor Subtypes is Not Mediated by Direct Protein-Protein Interaction Through Intracellular and Extracellular Regions

  • Kang, Yun-Kyung;Yoon, Tae-Sook;Lee, Kyung-Lim;Kim, Hwa-Jung
    • Archives of Pharmacal Research
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    • v.26 no.10
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    • pp.846-854
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    • 2003
  • The oligomerization of G-proteincoupled receptors (GPCRs) has been shown to occur by various mechanisms, such as via disulfide covalent linkages, non covalent (ionic, hydrophobic) interactions of the N-terminal, and/or transmembrane and/or intracellular domains. Interactions between GPCRs could involve an association between identical proteins (homomers) or non-identical proteins (heteromers), or between two monomers (to form dimers) or multiple monomers (to form oligomers). It is believed that muscarinic receptors may also be arranged into dimeric or oigomeric complexes, but no systematic experimental evidence exists concerning the direct physical interaction between receptor proteins as its mechanism. We undertook this study to determine whether muscarinic receptors form homomers or a heteromers by direct protein-protein interaction within the same or within different subtypes using a yeast two-hybrid system. Intracellular loops (i1, i2 and i3) and the C-terminal cytoplasmic tails (C) of human muscarinic (Hm) receptor subtypes, Hm1, Hm2 and Hm3, were cloned into the vectors (pB42AD and pLexA) of a two-hybrid system and examined for heteromeric or homodimeric interactions between the cytoplasmic domains. No physical interaction was observed between the intracellular domains of any of the Hm/Hm receptor sets tested. The results of our study suggest that the Hm1, Hm2 and Hm3 receptors do not form dimers or oligomers by interacting directly through either the hydrophilic intracellular domains or the C-terminal tail domains. To further investigate extracellular domain interactions, the N-terminus (N) and extracellular loops (o1 and o2) were also cloned into the two-hybrid vectors. Interactions of Hm2N with Hm2N, Hm2o1, Hm2o2, Hm3N, Hm3o1 or Hm3o2 were examined. The N-terminal domain of Hm2 was found to have no direct interaction with any extracellular domain. From our results, we excluded the possibility of a direct interaction between the muscarinic receptor subtypes (Hm1, Hm2 and Hm3) as a mechanism for homo- or hetero-meric dimerization/oligomerization. On the other hand, it remains a possibility that interaction may occur indirectly or require proper conformation or subunit formation or hydrophobic region involvement.

Beamforming Games with Quantized CSI in Two-user MISO ICs (두 유저 MISO 간섭 채널에서 불완전한 채널 정보에 기반한 빔포밍 게임)

  • Lee, Jung Hoon;Lee, Jin;Ryu, Jong Yeol
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.21 no.7
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    • pp.1299-1305
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    • 2017
  • In this paper, we consider a beamforming game between the transmitters in a two-user multiple-input single-output interference channel using limited feedback and investigate how each transmitter is able to find a modified strategy from the quantized channel state information (CSI). In the beamforming game, each of the transmitters (i.e., a player) tries to maximize the achievable rate (i.e., a payoff function) via a proper beamforming strategy. In our case, each transmitter's beamforming strategy is represented by a linear combining factor between the maximum ratio transmission (MRT) and the zero forcing (ZF) beamforming vectors, which is the Pareto optimal achieving strategy. With the quantized CSI, the transmitters' strategies may not be valid because of the quantization errors. We propose a modified solution, which takes into account the effects of the quantization errors.

A Novel Expression System for Recombinant Marine Mussel Adhesive Protein Mefp1 Using a Truncated OmpA Signal Peptide

  • Lee, Sang Jun;Han, Yun Hee;Nam, Bo Hye;Kim, Young Ok;Reeves, Peter R.
    • Molecules and Cells
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    • v.26 no.1
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    • pp.34-40
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    • 2008
  • To express an increased level of recombinant Mefp1 (marine mussel adhesive protein) in soluble form, we constructed expression vectors encoding truncated OmpA signal peptide-Mefp1 fusion proteins. OmpA signal peptide (OmpASP) is the 21 residue peptide fragment of the 23 residue OmpA signal sequence cleavable by signal peptidase I. We successfully produced increased levels of soluble recombinant Mefp1 (rMefp1) with various deletions of OmpASP, and found that the increased expression was caused by the increased pI of the N-terminus of the fusion proteins (${\geq}10.55$). All the OmpA signal peptide segments of 3-21 amino acids in length had the same pI value (10.55). Our results suggest that the pI value of the truncated OmpASP ($OmpASP_{tr}$) play an important role in directional signaling for the fusion protein, but we found no evidence for the presence of a secretion enhancer in OmpASP. For practical applications, we increased the expression of soluble rMefp1 with $OmpASP_{tr}$ peptides as directional signals, and obtained rMefp1 with the native amino terminus (nN-rMefp1) using an $OmpASP_{tr}$ Xa leader sequence that contains the recognition site for Xa protease.

A Study on Simultaneous Adjustment of GNSS Baseline Vectors and Terrestrial Measurements

  • Nguyen, Dinh Huy;Lee, Hungkyu;Yun, Seonghyeon
    • Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
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    • v.38 no.5
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    • pp.415-423
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    • 2020
  • GNSS (Global Navigation Satellite System) is mostly used for high-precise surveys due to its accuracy and efficiency. But this technique does not always fulfill the demanding accuracy in harsh operational environments such as urban canyon and forest. One of the remedies for overcoming this barrier is to compose a heterogeneous surveying network by adopting terrestrial measurements (i.e., distances and angles). Hence, this study dealt with the adjustment of heterogeneous surveying networks consisted of GNSS baseline vectors, distances, horizontal and vertical angles with a view to enhancing their accuracy and so as to derive an appropriate scheme of the measurement combination. Reviewing some technical issues of the network adjustments, the simulation, and experimental studies have been carried out, showing that the inclusion of the terrestrial measurements in the GNSS standalone overall increased the accuracy of the adjusted coordinates. Especially, if the distances, the horizontal angles, or both of them were simultaneously adjusted with GNSS baselines, the accuracy of the GNSS horizontal component was improved. Comparing the inclusion of the horizontal angles with those of the distances, the former has been more influential on accuracy than the latter even though the same number of measurements were employed in the network. On the other hand, results of the GNSS network adjustment together with the vertical angles demonstrated the enhancement of the vertical accuracy. As conclusion, this paper proposes a simultaneous adjustment of GNSS baselines and the terrestrial measurements for an effective scheme that overcomes the limitation of GNSS control surveys.