• Title/Summary/Keyword: 3' poly(A)-tail

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Post-transcriptional and translational regulation of mRNA-like long non-coding RNAs by microRNAs in early developmental stages of zebrafish embryos

  • Lee, Kyung-Tae;Nam, Jin-Wu
    • BMB Reports
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    • v.50 no.4
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    • pp.226-231
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    • 2017
  • At the post-transcriptional and translational levels, microRNA (miRNA) represses protein-coding genes via seed pairing to the 3' untranslated regions (UTRs) of mRNA. Although working models of miRNA-mediated gene silencing are successfully established using miRNA transfections and knockouts, the regulatory interaction between miRNA and long non-coding RNA (lncRNA) remain unknown. In particular, how the mRNA-resembling lncRNAs with 5' cap, 3' poly(A)-tail, or coding features, are regulated by miRNA is yet to be examined. We therefore investigated the functional interaction between miRNAs and lncRNAs with/without those features, in miRNA-transfected early zebrafish embryos. We observed that the greatest determinants of the miRNA-mediated silencing of lncRNAs were the 5' cap and 3' poly(A)-tails in lncRNAs, at both the post-transcriptional and translational levels. The lncRNAs confirmed to contain 5' cap, 3' poly(A)-tail, and the canonical miRNA target sites, were observed to be repressed in the level of both RNA and ribosome-protected fragment, while those with the miRNA target sites and without 5' cap and 3' poly(A)-tail, were not robustly repressed by miRNA introduction, thus suggesting a role as a miRNA-decoy.

A Study on the Mechanism of Photoluminescence in Poly(3-hexylthiophene) (Poly(3-hexylthiophene)의 PL 발광 메카니즘에 관한 연구)

  • 김주승;서부완;구할본
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.14 no.2
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    • pp.133-138
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    • 2001
  • We studied the optical properties of poly(3-hexylthiophene) for applying to the emitting material of organic electro luminescent device. The infrared spectrum and NMR of synthesized polymer gave good evidence for the conjugation of 3-hexylthiophene monomer unit. We confirmed that poly(3-hexylthiophene) contains the HT(head-to-tail)-HT(head-to-Tail) linkage larger than 65% based on NMR analysis. FTIR and raman spectroscopy show that poly(3-hexylthiophene) has two main vibration levels which have an energy about 0.18eV and 0.36eV. Electronic absorption spectra shifted to the shorter wavelength with increasing temperature, which is related to a conformational transition of the polymer. Photoluminescence spectrum generated at low temperature(10K) is separated at 669nm, 733nm and 812nm that it's because of phonon energy generated from the lattice vibration.

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Polyadenylation Is Dispensable for Encapsidation and Reverse Transcription of Hepatitis B viral Pregenomic RNA

  • Lee, Hye-Jin;Lee, Jehan;Shin, Myeong-Kyun;Ryu, Wang-Shick
    • Molecules and Cells
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    • v.25 no.4
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    • pp.545-552
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    • 2008
  • A hepadnaviruses replicates its DNA genome via reverse transcription of an RNA template (pregenomic RNA or pgRNA), which has a cap structure at the 5' end and a poly(A) tail at the 3' end. We have previously shown that the 5' cap is indispensable for encapsidation of the pgRNA. A speculative extension of the above finding is that the cap contributes to encapsidation via its interaction with the poly(A) tail, possibly involving eIF4E-eIF4G-PABP interaction. To test this hypothesis, poly(A)-less pgRNAs were generated via cleavage by a cis-acting hepatitis delta virus ribozyme sequence. We found that accumulation of the poly(A)-less pgRNA was markedly diminished, mostly likely due to its reduced stability. Importantly, however, the remaining poly(A)-less pgRNAs were nonetheless encapsidated and reverse transcribed normally when the reduced stability was taken account. Our finding clearly contradicts the notion that the poly(A) tail has any function in encapsidation and viral reverse transcription.

Electrochemical Behavior of Poly 8-(3-Acetylimino-6-methyl 2,4-dioxopyran)-1-aminonaphthaline in Aqueous and Non Aqueous Media

  • Hathoot, A.A.
    • Bulletin of the Korean Chemical Society
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    • v.24 no.11
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    • pp.1609-1612
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    • 2003
  • The electrooxidation of 8-(3-acetylimino-6-methyl 2,4-dioxopyran)-1-aminonaphthaline (AMDAN) in aqueous and non aqueous media led to the formation of polymeric films, poly (AMDAN). The monomer, undergo anodic oxidation through the formation of a monocation radical irrespective of the nature of the medium. In aqueous medium, the monocation radical undergoes, through its resonance structures, dimerisation involving tail-to-tail, head-to-tail and even head-to-head coupling. The products formed, being more easily oxidisable than the parent substance, undergo further oxidation at the same potential so that the overall oxidation involves a one-step (i.e., a single wave), two-electron process. In non-aqueous medium, the monocation radical does not undergo dimerisation through coupling reactions. Retaining its identity, monomer oxidise in two steps involving one electron in each step. The fact that the cathodic peaks corresponding to these anodic peaks are rarely observed indicates fast consumption of the electrogenerated monocation radicals and dications by follow-up chemical reactions to produce polymeric products (poly AMDAN). The electrochemical behavior of the formed polymer films was investigated in both non aqueous and aqueous media. The films prepared in non aqueous medium were found to be more electroactive than that the films prepared in aqueous medium. This is confirmed with the results in litreature which illustrate that the film prepared in aqueous solution hold water in its structure via hydrogen bonding, which causes decomposition reactions.

Immunological Detection of Garlic Latent Virus (마늘 잠복 바이러스의 면역학적 진단)

  • Choi, Jin-Nam;Song, Jong-Tae;Song, Sang-Ik;Ahn, Ji-Hoon;Choi, Yang-Do;Lee, Jong-Seob
    • Applied Biological Chemistry
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    • v.38 no.1
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    • pp.49-54
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    • 1995
  • To understand the molecular structure and pathogenesis mechanism of Korean garlic viruses, we have isolated cDNA clones for garlic viruses. The partial nucleotide sequences of 24 cDNA clones were determined and those of five clones containing poly(A) tail were compared with sequences of other plant viruses. One of these clones, V9, has a primary structure similar to the carlavirus group, suggesting that the clone V9 derived from a part of garlic latent virus (GLV). Northern blot analysis with the clone V9 as a probe demonstrated that GLV genome is 8.5 knt long and has a poly(A) tail. The clone V9 encodes coat protein (CP) of 33 kDa and nucleic acid binding protein of 10 kDa in different reading frame. The hexanucleotide motif, 5'-ACCUAA, which is conserved in the 3' noncoding region arid was proposed to be a cis-acting element involved in the production of negative strand genomic RNA was noticed. Complementary sequence to the hexanucleotide motif, 5'-TTAGGT, is also found in the positive strand of V9 RNA. The putative CP gene was cloned into the pRSET-A expression vector and expressed in E. coli BL21. The expressed recombinant V9CP protein was purified by $Ni^{2+}$ NTA affinity chromatography. The anti-V9CP antibody recognizes 34 kDa polypeptide which could be CP of GLV in infected garlic leaf extract. Immunoblot and Northern blot analysis of various cultivars shows wide occurrence of GLV in Korean garlic plants.

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Characteristics of Electronic Absorption Spectrum and Photoluminescence in Cast-Poly(3-hexylthiophene) Films (캐스팅법으로 제작한 Poly(3-hexylthiophene)의 흡수스펙트럼에 따른 형광 특성)

  • 김주승;구할본;조재철
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 1998.11a
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    • pp.57-60
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    • 1998
  • Poly(3-hexylthiophene)(P3HT) was synthesized by use of FeCl$_3$ as a oxidizing agent at $25^{\circ}C$. The infrared spectrum of our polymer gave good evidence for the conjugation of 3-hexylthiophene monomer unit. P3HT contains the HT(head-to-tail) linkage larger than 64% based on NMR analysis. Electronic absorption and photoluminescence studies show that cast films of P3HT have three exciting state. Absorption spectrum was separated with three maximum peaks by Giese-French method and shifted to the shorter wavelength with increasing temperature. Separated absorption spectrum of P3HT is well adapted to PL peak appeared at longer wavelength. Low temperature PL spectrum is well separated at 669nm, 733nm and 812nm.

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Cloning and Nucleotide Sequence of a cDNA Encoding the Rat Triosephosphate Isomerase

  • Lee, Kyunglim;Ryu, Jiwon
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.497-501
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    • 1996
  • A gene coding for triosephosphate isomerase (TPI) from a rat skeletal muscle cDNA library was cloned and its nucleotide sequence was determined. The 1, 348-bp cDNA clone contains 24 bp $5^I$ noncoding region, the entire 750 bp coding region corresponding to a protein of 249 amino acids, $547bp 3^I$ noncoding region and part of a poly(A) tail. It also contains a polyadenylation signal, AATAAA, starting from 17 bp upstream of the poly(A) tail. The calculated molecular weight of rat TPI is 27.8 kDa and the net charge is +4. The deduced amino acid sequence from rat TPI CDNA sequence has 93% and 94% homology with that of mouse and human clones, respectively. The amino acids at the residue of Asn12, Lys14, His96, Glu 166, His96, His101, Ala177, Tyr165, Glu13O, Tyr2O9, and Ser212 in catalytic site are completely identical, confirming that the functional residues in TPI proteins are highly conserved throughout evolution. The most profound characteristic of rat TPI enzyme, compared with other TPIs, is that there are five cysteine substitutions at the residue of 21, 27, 159, 195 and 204. A Glu123 instead of Gly was found in rabbit, rhesus, mouse and human sequences. Through the method of RT-PCR, the mRNA transcription level of TPI gene was found to be different among various tissues and was highest in muscle.

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Nucleotide Sequence of Leghemoglobin cDNA from Canavalia lineata

  • An, Chung-Sun
    • Journal of Plant Biology
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    • v.37 no.2
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    • pp.167-173
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    • 1994
  • Poly(A)+ RNA was selected from Canavalia lineata root nodule RNA through oligo(dT) cellulose column and used for construction of a cDNA library using λgt10-EcoRI arms. The size of the library was 7.2$\times$105 pfu/mL. A full length leghemoglobin (Lb) cDNA clone, pCILb1(687 bp) isolated with soybean Lb probe, contained one open reading frame (ORF) of 447 bp with 54 bp plus 186 bp at 5' and 3' untranslated region, respectively. A consensus sequence of plant translation start region (AAAATGGG) was found at 5' untranslated region, and two polyadenylation-related sequence (AATAAA, AATAAG) and a conserved motif between them (gACTTGTT) were found upstream of poly(A)+ tail consisted of 13 (A)s at 3' untranslated region. The ORF encoded a polypeptide consisted of 149 amino acids with a molecular weight of 16.2 kD. Deduced amino acid sequences showed high degree of homology values with those of other Lbs ranging from 66% (Casuarina glauca) to 85% (Glycine max).

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Molecular Cloning of the 3'-Terminal Region of Garlic Potyviruses and Immunological Detection of Their Coat Proteins

  • Song, Sang-Ik;Song, Jong-Tae;Chang, Moo-Ung;Lee, Jong-Seob;Park, Yang-Do
    • The Plant Pathology Journal
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    • v.15 no.5
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    • pp.270-279
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    • 1999
  • cDNAs complementary to the 3'-terminal regions of two potyvirus genomes were cloned and sequenced. The clone G7 contains one open reading frame (ORF) of 1,338 nucleotides and a 3' untranslated region (3'-UTR) of 403 nucleotides at the 3'-end excluding the 3'end poly(A) tail. The putative viral coat protein (CP) shows 55%-92% amino acid sequence homology to those of Allium potyviruses. The genome size of the virus was analyzed to be about 9.0 kb by Northern blot analysis. Five cDNA clones were screened out using GPV2 oligonucleotide as a probe. One of these clones, DEA72, which has a longest cDNA insert, contains one ORF of 1,459 nucleotides and a 3'-UTR of 590 nucleotides at the 3'-end excluding the 3'-end poly(A) tail. The putative viral CP shows 57%-88% amino acid sequence homologies to those of Allium potyviruses. The genome size of the virus was analyzed to be about 9.6 kb by Northern blot analysis. The results of immunoblot and Northern blot analyses suggest that almost all of the tested garlic plants showing mosaic or streak symptoms are infected with DEA72-potyvirus in variable degrees but rarely infected with G7-potyvirus in variable degrees but rarely infected with DEA72-potyvirus in variable degrees but rarely infected with G7-potyvirus. Immunoelectron microscopy using anti-DEA72 CP antibody shows that this potyvirus is about 750 nm long and flexuous rod shaped.

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Characterization of Mud Loach (Misgurnus mizolepis) Apolipoprotein A-I: cDNA Cloning, Molecular Phylogeny and Expression Analysis (미꾸라지(Misgurnus mizolepis) Apolipoprotein A-I cDNA의 구조, 분자계통 및 발현 특징 분석)

  • Lee, Youn-Ho;Noh, Jae-Koo;Kim, Keun-Yong;Cho, Young-Sun;Nam, Yoon-Kwon;Kim, Dong-Soo
    • Journal of Aquaculture
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    • v.20 no.1
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    • pp.65-72
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    • 2007
  • Full length complementary DNA encoding apolipoprotein A-I (apoA-I) was isolated and characterized in mud loach (Misgurnus mizolepis). Mud loach apoA-I cDNA encoding 24 bp of 5'-untranslated region (UTR), 762 bp of single open reading frame (ORF) consists of 254 amino acids and 293 bp of 3'-UTR excluding stop codon and poly (A+) tail. Two overlapping polyadenylation signals (AATAAAATAAA) was found 9 bp prior to the poly (A+) tail. Mud loach apoA-I represented considerable homology to those from other teleost species at amino acid level with conserving common features of vertebrate apoA-I. Molecular phylogenetic analysis inferred the phylogenetic hypothesis that was generally in accordance with the previous taxonomic relationship. Apolipoprotein A-I mRNA was detected in various tissues, but the mRNA levels were quite varied depending on tissues based on semi-quantitative RT-PCR. Liver and brain showed the significantly higher levels of apoA-I transcripts than other tissues. mRNA expression of apoA-I was quite low in very early stage of embryonic development, however dramatically enhanced from 8 hours post fertilization. This increased mRNA level was retained consistently up to 14 days post hatching.