• Title/Summary/Keyword: oligonucleotides

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Transgelin is Required for Agonist-induced $Ca^{2+}$-Sensitization in Vascular Contractility: Evidence from an Antisense Approach (칼슘이온 감작이 포함된 Transgelin의 혈관 평활근 수축성 조절)

  • Je, Hyun-Gon;Je, Hyun-Dong
    • YAKHAK HOEJI
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    • v.53 no.3
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    • pp.156-160
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    • 2009
  • The present study was undertaken to determine whether transgelin participates in the regulation of vascular smooth muscle contraction and, if so, to investigate the mechanism. By PCR homology cloning, the cDNA sequence of ferret transgelin was determined and phosphorothioate antisense and random oligonucleotides were synthesized and introduced into strips of ferret aorta by a chemical loading procedure. Treatment of ferret aorta with transgelin antisense oligonucleotides resulted in a significant decrease in protein levels of transgelin to sham- or random sequence-loaded muscles, but no change in the protein levels of actin. Contraction in response to a phorbol ester was significantly decreased in antisense-treated muscles compared to sham- or random sequence-loaded controls. Neither basal intrinsic tone nor the contraction in response to phenylephrine was significantly affected by the antisense treatment. The data indicate that transgelin plays a significant role in the regulation of contraction and suggest that in a tonically active smooth muscle transgelin may function as a signalling protein to facilitate PKC or ERK-dependent signalling rather than thick filament regulation including $Ca^{2+}$ or calmodulin dependent regulation of myosin light chain kinase.

Effect of Benzalkonium Chloride on Percutaneous Absoption of Antisense Phosphorothioate Oligonucleotides

  • Lee, Young-Mi;Lee, Sung-Hee;Ko, Geon-Il;Kim, Jae-Baek;Sohn, Dong-Hwan
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.435-440
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    • 1996
  • The effect of benzalkonium chloride on skin permeability of partially modified antisense phosphorothioate oligonucleotides (PS-ODN), which are designed as scar formation inhibitor, was investigated using Franz Diffusion Cell. When the concentration ratio of PS-ODN-quarternary ammonium salt complex is more than 1:100, the apparent partition coefficient (APC) of each complex was increased in the following order; tetraphenyl phosphonium chloride (TPP) < cetyltrimethyl ammonium bromide(CTAB) < benzalkonium chloride (BZ). The permeability of PS-ODN through the rat skin increased in the presence of BZ. The fluxs of PS-ODN with BZ were increased by addition of Pluronic F 68 or Triton X-100 to phosphate buffered saline (PBS), respectively. When the mole ratio of PS-ODN to BZ is 1:10, the fluxs penetrated of PS-ODN with BZ was greatest. The increase of the permeability in the presence of BZ might be due to the formation of lipophilic ion-pair complex between PS-ODN and BZ. By regulation of mole ratio of PS-ODN to BZ, the development of topical dosage forms using PS-ODN as scar formation inhibitor will be possible with minimal systemic exposure.

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Real-Time Detection of DNA Hybridization Assay by Using Evanescent Field Microscopy

  • Kim, Do-Kyun;Choi, Yong-Sung;Murakami, Yuji;Tamiya, Eiichi;Kwon, Young-Soo
    • KIEE International Transactions on Electrophysics and Applications
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    • v.11C no.3
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    • pp.85-90
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    • 2001
  • The determination of DNA hybridization reaction can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and other application area. So, the improvement of DNA detection system is very important for the determination of this hybridization reaction. In this study, we report the characterization of the probe and target oligonucleotide hybridization reaction using the evanescent field microscopy. First, we have fabricated DNA chip microarray. The particles which were immobilized oligonucleotides were arranged by the random fluidic self-assembly on the pattern chips, using hydrophobic interaction. Second, we have detected DNA hybridization reaction using evanescent field microscopy. The 5'-biotinylated probe oligonucleotides were immobilized on the surface of DNA chip microarray and the hybridization reaction with the Rhodamine conjugated target oligonucleotide was excited fluorescence generated on the evanescent field microscopy. In the foundation of this result, we could be employed as the basis of a probe olidonucleotide, capable of detecting the target oligonucleotide and monitoring it in a large analyte concentration range and various mismatching condition.

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Detection and Analysis of DNA Hybridization Characteristics by using Thermodynamic Method (열역학법을 이용한 DNA hybridization 특성 검출 및 해석)

  • Kim, Do-Gyun;Gwon, Yeong-Su
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.51 no.6
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    • pp.265-270
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    • 2002
  • The determination of DNA hybridization reaction can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and application area. So, the improvement of DNA hybridization detection method is very important for the determination of this hybridization reaction. Several molecular biological techniques require accurate predictions of matched versus mismatched hybridization thermodynamics, such as PCR, sequencing by hybridization, gene diagnostics and antisense oligonucleotide probes. In addition, recent developments of oligonucleotide chip arrays as means for biochemical assays and DNA sequencing requires accurate knowledge of hybridization thermodynamics and population ratios at matched and mismatched target sites. In this study, we report the characteristics of the probe and matched, mismatched target oligonucleotide hybridization reaction using thermodynamic method. Thermodynamic of 5 oligonucleotides with central and terminal mismatch sequences were obtained by measured UV-absorbance as a function of temperature. The data show that the nearest-neighbor base-pair model is adequate for predicting thermodynamics of oligonucleotides with average deviations for $\Delta$H$^{0}$ , $\Delta$S$^{0}$ , $\Delta$G$_{37}$ $^{0}$ and T$_{m}$, respectively.>$^{0}$ and T$_{m}$, respectively.

Genetic Distances Within-Population and Between-Population of Tonguesole, Cynoglossus spp. Identified by PCR Technique

  • Yoon, Jong-Man
    • Development and Reproduction
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    • v.23 no.3
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    • pp.297-304
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    • 2019
  • The higher fragment sizes (>2,100 bp) are not observed in the two C. spp. populations. The six oligonucleotides primers OPA-11, OPB-09, OPB-14, OPB-20, OPC-14, and OPC-18 were used to generate the unique shared loci to each tonguesole population and shared loci by the two tonguesole populations. The hierarchical polar dendrogram indicates two main clusters: Gunsan (GUNSAN 01-GUNSAN 11) and the Atlantic (ATLANTIC 12-ATLANTIC 22) from two geographic populations of tonguesoles. The shortest genetic distance displaying significant molecular difference was between individuals' GUNSAN no. 02-GUNSAN no. 01 (genetic distance=0.038). In the long run, individual no. 02 of the ATLANTIC tonguesole was most distantly related to GUNSAN no. 06 (genetic distance=0.958). These results demonstrate that the Gunsan tonguesole population is genetically different from the Atlantic tonguesole population. The potential of PCR analysis to identify diagnostic markers for the identification of two tonguesole populations has been demonstrated. As a rule, using various oligonucleotides primers, this PCR method has been applied to identify polymorphic/specific markers particular to species and geographical population, as well as genetic diversity/polymorphism in diverse species of organisms.

PCR Analysis for Genetic Distances of Two Charybdis Crab Populations

  • Yoon, Jong-Man
    • Development and Reproduction
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    • v.26 no.2
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    • pp.91-98
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    • 2022
  • Genomic DNA (gDNA) set apart from two populations of Korean Charybdis crab (Charybdis japonica) was augmented by PCR experiments. The five oligonucleotides primers (ONT-primers) were spent to yield the number of unique loci shared to each crab population (ULSECP) and number of loci shared by the two crab populations (LSTCP). 305 fragments (FRAGs) were identified in the Charybdis crab population A (CCPA), and 344 in the Charybdis crab population B (CCPB): 44 number of ULSECP (14.43%) in the CCPA and 110 (31.98%) in the CCPB. 44 number of LSTCP, with an average of 8.8 per primer, were detected in the two crab populations. The bandsharing (BS) value between entity's no. 01 and no. 10 was the lowest (0.371) between the two CCPs. The average bandsharing (ABS) values of individuals in the CCPA (0.575±0.014) were lesser than in those originated from the CCPB (0.705±0.011) (p < 0.05). The polar hierarchical dendrogram (PHD) achieved by the five ONT-primers denotes three genetic clusters (GCs): cluster I (CHARYBCRAB 01, 04, 05, 06, and 08), cluster II (CHARYBCRAB 02, 03, 07, 09, 10, and 11) and cluster III (CHARYBCRAB 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, and 22). The shortest genetic distance (GD) displaying significant molecular difference (MD) was between individuals CHARYBCRAB no. 18 and CHARYBCRAB no. 17 (0.055).

Genetic Distances of Binary Pen Shell Atrina pectinata Populations

  • Yoon, Jong-Man
    • Development and Reproduction
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    • v.26 no.3
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    • pp.127-133
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    • 2022
  • The seven oligonucleotides primers were consumed to produce the quantity of unique loci shared to each pen shell team (ULSEPT) and quantity of loci shared by the binary pen shell teams. 154 quantities of LSBPP, with a mediocre of 22.0 per primer, were noticed in the binary pen shell (Atrina pectinata) teams. 328 fragments were recognized in the pen shell team A (PSTA), and 257 in the pen shell team B (PSTB): 77 quantities of ULSEPT (23.48%) in the PSTA and 121 (47.08%) in the PSTB. The band-sharing amount (BS amount) between entity's no. 01 and no. 05 was the highest (0.884) between the binary PSTs. The median band-sharing amount of entities in the PSTA (0.685±0.011) was higher than in those invented from the PSTB (0.640±0.009) (p<0.05). The highest genetic distance presenting substantial molecular difference was between entities PECTINATA no. 06 and PECTINATA no. 04 (0.498). Through this study, it is possible a certain degree to contribute to increasing the cultivation of pen shells, conservation of species, protection of the natural environment, and preservation of ecosystems.

Genetic Distances for Intra- and Between-Group of Scapharca subcrenata from Yeosu of the Korea

  • Yoon, Jong-Man
    • Development and Reproduction
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    • v.25 no.4
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    • pp.305-311
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    • 2021
  • The oligonucleotides polymers (ON-polymers) were used producing a total of 110 loci unique to each clam population (LUECP) in group one and 132 in group two, respectively, varying in amount of DNA fragments (FRs) from greater than near 50 to a smaller quantity than 1,050 bp. The larger FR amounts (>1,050 bp) are not noticed in the two Scapharca subcrenata groups. The ON-polymer OPD-01 produced 33 LUECP, which were defining each group, almost 300 bp, 450 bp, and 500 bp, in the group one. The OPD-15 recognized 22 loci shared by the two clam populations (Loci shared by the two clam populations, LSTCP), a variety of FRs of sizes 300 bp that were equivalent in all specimens. The mean number of LUECP was varied and 1.2-fold greater in the shellfish group two than in the group one. Respecting mean bandsharing (BS) grade outcomes, entities in the shellfish group one (0.779±0.011) had a little higher BS grades than did entities from the group two (0.756±0.009) (p<0.05). The entities of the shellfish group one are not tightly gathered with other entities of the group two. The genetic distance (GD) (0.422) of this invertebrate (SUBCRENATA 02 and 01) is 7.41-fold hereditarily distinct to the GD (0.057) of the other invertebrate (SUBCRENATA 22 and 19). The polar dendrogram (PDG) procured by the five ON-polymers underlines two characteristic groups.

Molecular Cloning and Nucleotide Sequencing of a DNA Clone Encoding Arginine Decarboxylase in Rice (Oryza sativa L.) (벼의 arginine decarboxylase DNA clone의 재조합 및 염기서열 분석)

  • Hong, Sung-Hoi;Jeung, Ji-Ung;Ok, Sung-Han;Shin, Jeong-Sheop
    • Applied Biological Chemistry
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    • v.39 no.2
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    • pp.112-117
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    • 1996
  • Arginine decarboxylase (ADC) is the first enzyme in one of the two pathways of diamine putrescine biosynthesis in plants. The genes encoding ADC have previously been cloned from Escherichia coli, oat and tomato genome. Two degenerate oligonucleotides (17-mer) corresponding to two conserved regions of ADC were used as primers in polymerase chain reaction of rice (Oryza sativa L.) genomic DNA, and an approximately 1.0 kbp fragment was obtained. This amplified PCR product showed an open reading frame which contains 1,022 bp of nucleotide sequences. This PCR product was cloned into pGEM-originated T vector and the short 500 bp PstI digested fragment was subcloned into pGEM-3zf(+/-) vectors to facilitate sequencing. The nucleotide sequence of this PCR product showed about 74% and 70% identity with the same regions of the oat and tomato ADC cDNA sequences, respectively. The predicted amino acid sequence exhibited 45% and 62% identity with oat and tomato ADC polypeptide fragments, respectively. The sequence similarities of 34%, 47% and 38% were previously reported in oat and E. coli, tomato and oat, and tomato and E. coli ADC amino acids, respectively. Therefore, similarities and identities between rice and oat or tomato are remarkably higher than those others of the previous reports. In the highly conserved regions in both the amino acid sequence and spacing regions among the sequences of these three, rice ADC open reading frame also has the exactly same regions with the striking similarity. RNA blot analysis showed that hnc is expressed as a transcript of approximately 2.5 kbP in the rice seedling leaf tissues.

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