• Title/Summary/Keyword: Polyadenylation

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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 and Characterization of the Gene Encoding Chitinase from Bombyx mandarina (멧누에(Bombyx mandarina)로부터 Chitinase를 코딩하는 cDNA의 분리 및 염기서열 결정)

  • 구태원;황재삼;성규병;윤은영;방혜선;권오유
    • Journal of Life Science
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    • v.9 no.4
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    • pp.341-347
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    • 1999
  • Insects use chitinolytic enzyme to digest chitin in the exoskelton during the molting process. We have isolated and sequenced a chitinase-encoding cDNA from the silkworm, Bombyx mandarina, compared its sequenced with genes encoding chitinolytic enzymes from other sources. The insert DNA in the clone is 2,675 nucleotides long with an open reading frame of 1,695 uncletides that encodes a protein of 565 amino acids with a molecuar weight of 63.4 kDa. The 3' -untranslated region of 889 nucleotides is AT-rich and contains two putative polyadenylation signals. The N-terminal sequence of the encoded protein contains numerous hydrophobic residues characteristic of a leader peptide. The amino acid alignment revealed that the endo-$\beta$-N-acetylglucosaminidase had 83% and 97% homology with M. sexta and B. mori, respectively. The deduced amino acid had two highly conserved region at the amino acid residues 97-111 and 139-148 that were related to the existing chitinase.

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Frequent Changes of 3' UTR Sequences in the Genes Expressed During Hematopoietic Differentiation Implicates the Importance of 3' UTR in Regulation of Gene Function (조혈세포의 분화과정에서 발현되는 유전자의 3‘ UTR 염기서열의 변화가 유전자 기능의 조절에 미치는 영향에 대한 연구)

  • Lee Sanggyu
    • YAKHAK HOEJI
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    • v.49 no.3
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    • pp.205-211
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    • 2005
  • The 3' UTR (3' untranslated region) plays important roles in controlling gene expression through regulating 3' polyadenylation, mRNA export, subcellular localization, translational efficiency, and mRNA stability. Changes in the 3' UTR sequence in an expressed transcript can result in functional changes of the genes that are expressed in pathological conditions compared with those genes expressed in normal physiologic conditions. A genome-wide survey of 3' UTR variation was performed for the genes expressed during hematopoietic differentiation from CD34+ stem/progenitor cells to CD 15 + myeloid progenitor cells. Wide-spread differential usage of the 3' UTR was observed from the genes expressed during this cellular transition. This study implies that the 3' UTR can be a highly coordinated region for post-transcriptional regulation of the function of expressed genes.

Protein-Protein Interaction between Poly(A) Polymerase and Cyclophilin A in Chemotactic Cells

  • Choi, Hyun-Sook;Kim, Hana;Lee, Changgook;Kim, Youngmi;Lee, Younghoon
    • Bulletin of the Korean Chemical Society
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    • v.35 no.1
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    • pp.83-86
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    • 2014
  • Poly(A) polymerase (PAP) play an essential role for maturation of mRNA by adding the adenylate residues at the 3' end. PAP functions are regulated through protein-protein interaction at its C-terminal region. In this study, cyclophilin A (CypA), a member of the peptidyl-prolyl cis-trans isomerase family, was identified as a partner protein interacting with the C-terminal region PAP. The interaction between PAP and CypA was inhibited by the immunosuppressive drug cyclosporine A. Deletion analysis revealed that the N-terminal 56 residues of CypA are sufficient for the interaction with PAP. Interestingly, we observed that PAP and CypA colocalize in the nucleus during SDF-1-induced chemotaxis, implying that CypA could be involved in the regulation of polyadenylation by PAP in the chemotactic cells.

Molecular Characterization of Plant Genes (식물 유전자의 구조와 특성)

  • 이종섭
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.19-49
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    • 1987
  • Recent development of recombinant DNA techniques such as gene cloning and DNA sequencing has led to understanding of genetic information coded on plant genes and their application to crop improvements. Nuclear genes so far isolated and characterized at the molecular level from various plants are those involved mainly in photosynthesis, nitrogen fixation, seed development and defensive responses to environmental stresses. Most of plant genes contain intervening sequences (introns) flanked with GT and AG, as it typical of animal genes. The 5' flanking regions of plant gene revealed the presence of promoter elements such as TATAAA and CCAAT, which have been identified at animal genes to be involved in transcrip- tion initiation. The 3' untranslated regions include a sequence similar to AATAAA whcih functions as a polyadenylation signal in other eukaryotic genes. Furthermore, enhancer-type sequences were found at the 5' flanking regions of various plant genes. This indicates that the structure of plant genes is very similar to animal genes and mechanisms governing the synthesis and processing of mRNAs may be identical in higher eukaryotes. However, genes expression studies involving transformation revealed their differ ences within plants and between plant and animal systems.

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Studies on Peroxidase Isozymes of Tobacco and Korean Radish: implication of Specific Isoperoxidase in Shoot Formation (담배와 무 Peroxidase Isozyme들의 세포 생화학적 연구: 조직 특이성 발현과의 연관성)

  • 김승수
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.357-389
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    • 1987
  • In an attempt to clarify the physiological functions of individual isoperoxidases, we have studied enzymatic and immunological properties as well as cellular distribution of isoperoxidases from tobacco callus and Korean radish. The gene expression patterns of isoperoxidases in shoot and non-shoot-forming tobbaco callus were also examined by rabbit reticulocyte lysatein vitro translation system. These results indicate that fraction of translatable poly(A)-isoperoxidase mRNA was increased considerably in shoots. At the present time, at least 6-7 isoperoxidases could be detected from the translation mixture of total cellular RNA, among which only one cell wall localized anodic isoperoxidase (named A3) mRNA was bimorphic mRNA. These data suggest the possible regulation of peroxidase activity during shoot formation by altering the polyadenylation state of mRNA. In case of Korean radish seedlings, poly(A)- peroxidase mRNA were also increased depending upon aging.

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A Study on the Transmission of a Transgene in the Offspring of Transgenic Mice (형질전환 생쥐의 후손에서 외래 유전자의 유전성에 대한 연구)

  • 염행철
    • Korean Journal of Animal Reproduction
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    • v.20 no.4
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    • pp.453-458
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    • 1997
  • It is known that the incorporation of genes into transgenic mice is generally stable and is p passed on to succeeding generations in a Mendelian fashion. In this report, transgenic mice were set as a model to evaluate whether the transgenes are transmitted in a Mendelian principle in a successive generations and how they are tran s smitted into their offspring. A 3.0 kb linear DNA fragment, containing the MMTV LTR, bovine aSI casein cDNA and SV 40 splicing and polyadenylation site; was microinjected into fertilized mouse embryos. The tail DNAs of the resulting pups were subjected to dot and Southern hybridizations to screen transgenic founders. The DNAs of their offspring were anlyzed by PCR to confirm the transmission of the transgene from F0. Out of 72 live pups four pups (5.6%), 3 males and 1 female, were positive for the transgene. The rates of transmission from F0 into F1 were 33.3, 7.7, 0, and 62.5%. Those from F1 into F2 were 63.6, 5.9, and 68.8% and those from F2 into F3 were 85.7, and 88.2%. In this report, the transmission pattern of transgenes in transgenic mice into their offspring was demonstrated. It either follows or does not follow in a Mendelian fashion. Deletion or loss of the transgenes from F0 in some lines became apparant to the succeeding generations.

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The Biological Functions of Plant Long Noncoding RNAs (식물의 긴비암호화 RNA들의 생물학적 기능)

  • Kim, Jee Hye;Heo, Jae Bok
    • Journal of Life Science
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    • v.26 no.9
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    • pp.1097-1104
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    • 2016
  • With the development of next generation sequencing (NGS), large numbers of transcriptional molecules have been discovered. Most transcripts are non -coding RNAs (ncRNAs). Among them, long non-coding RNAs (lncRNAs) with more than 200 nucleotides represent functional RNA molecule that will not be translated into protein. In plants, lncRNAs are transcribed by RNA polymerase II (Pol II) or Pol III, Pol VI and Pol V. After transcription of these lncRNAs, more RNA processing mechanisms such as splicing and polyadenylation occurs. The expression of plant lncRNAs is very low and is tissue specific. However, these lncRNAs are strongly induced by specific external stimuli. Because different external stimuli including environmental stresses induce a large number of plant lncRNAs, these lncRNAs have been gradually considered as new regulatory factors of various biological and development processes such as epigenetic repression, chromatin modification, target mimicry, photomorphogenesis, protein relocalization, environmental stress response, pathogen infection in plants. Moreover, some lncRNAs act as precursor of short RNAs. Although a large number of lncRNAs have been predicted and identified in plants, our current understanding of the biological function of these lncRNAs is still limited and their detailed regulatory mechanisms should be elucidated continuously. Here, we reviewed the biogenesis and regulation mechanisms of lncRNAs and summarized the molecular functions unraveled in plants.

The Structure and The Reason for Nuclear Accumulation of Poly A(-) Spliced SV40 RNA (Poly A tail이 없는 SV 40 spliced RNA의 구조 및 핵내 축적의 원인)

  • 박주상;노정혜
    • Korean Journal of Microbiology
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    • v.27 no.1
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    • pp.1-9
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    • 1989
  • The locations of 5' ends as well as the splicing pattern of viral poly A(-) 19S RNA from monkey cells infected with SV40 were determined by a modification of primer extension method. The 5' end of this RNA mapped at the major cap site at nucleotide residue 325, used most frequently by SV40 late RNAs. The intron from nt.373 to nt.558 was removed as the ordinary cytoplasmic poly A(+) 19S RNA. The 3'end of this RNA was very heterogeneous and distributed over 1 kb upstream of polyadenylation site, as determined by S1 nuclease mapping. The reason for this normally initiated and spliced RNA to accumulate in the nucleus was investigated. In order to test whether the presence of unused 3' splice region on this RNA caused such subcellular distribution, cells were transfected with SV40 mutant KNA containing deletion around 3' splice site. The RNA deleted of 3' splice region accumulated mainly in the cytoplasm. This accumulation did not result from the increased stability of the RNA due to the deletion, since the wild type and mutant RNAs exhibited similar half lives after chase with actinomycin D. Therefore it is likely that the 19S spliced RNA is hindered from being transported into the cytoplasm due to some pre-splicing complexes formed at the unused 3' splice site.

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Effect of Estrogen on the Gestational Profiles in Gene Expression of Placental Lactogen I, II and Pit-i in the Rat Placenta (흰쥐 태반에서 Placental Lactogen I과 II 그리고 Pit-1의 유전자 발현에 미치는 에스트로겐의 영향)

  • 정진권;강성구;강해묵;이병주
    • The Korean Journal of Zoology
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    • v.39 no.1
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    • pp.115-121
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    • 1996
  • To investigate gestational profiles in gene expression of placental lactogen I fpL4), PL-lI and Pit-1, RNA samples were extracted from the placentas of pregnant day 12 to 20 at 2 day intervals. Northern blots showed changes in gene expression of PL4, - 11 and Pit-i. Sizes of PL-l and -II mRNA were changed and amounts of PL-I, -H and Pit-1 mRNA increased during progress of gestation. To examine the effect of estrogen on the gene expression of PL-I, -Il and Pit-1, pregnant female rats were ovariectomized (OVX) and daily injected with estradiol (OVX + E). OVX markedly lowered the amount of PL4 and 41 mRNA, and shifted niRNA size from 1 kb to i 3 kb in PL-l mRNA and 0.6 kb to i kb in PL-ll mRNA, respectively. OVX had no effect on the mRNA size of Pit-1, but markedly attenuated Pit-1 mRNA level. Estrogen injection reversed the effect of OVX on the size-shift but not on the amount of PL4 and -Il mRNA. Replacement of E partially recovered OVX-induced inhibition of Pit-i mRNA level. Present results suggest that estrogen may play a pivotal role on the gene expression of PL-l and -Il such as alternative RNA splicing and/or polyadenylation, and Pit-1 may be involved in the gene expression of PL-l and 41 by estrogen.

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