• Title/Summary/Keyword: DNA/RNA bases

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DEGRADATION OF NUCLEIC ACIDS BY CELL-FREE EXTRACT OF MIXED RUMEN PROTOZOA OF BUFFALO RUMEN

  • Sinha, P.R.;Dutta, S.M..
    • Asian-Australasian Journal of Animal Sciences
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    • v.1 no.4
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    • pp.219-222
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    • 1988
  • Degradation of deoxyribonucleic acid(DNA) and ribonucleic acid(RNA) by cell-free extract of mixed rumen protozoa of buffalo rumen was investigated. DNA was observed to be degraded rapidly during an initial incubation period of 2 hr with simultaneous appearance of degradation products. RNA on the other hand recorded a rapid degradation during an initial incubation period of 1 hr. RNA degradation products appeared upto an incubation period of 2 hr. DNA was observed to degrade into oligo- and mononucleotides. pyrimidine nucleosides, purine nucleoside adenosine and bases xanthine, hypoxanthine and thymine. Degradation products of RNA comprised of pyrimidine nucleosides, purine nucleoside, adenosine and bases xanthine, hypoxanthine and uracil besides oligo- and mononucleotides.

Editing of Genomic TNFSF9 by CRISPR-Cas9 Can Be Followed by Re-Editing of Its Transcript

  • Lee, Hyeon-Woo
    • Molecules and Cells
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    • v.41 no.10
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    • pp.917-922
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    • 2018
  • The CRISPR-Cas system is a well-established RNA-guided DNA editing technique widely used to modify genomic DNA sequences. I used the CRISPR-Cas9 system to change the second and third nucleotides of the triplet $T{\underline{CT}}$ of human TNSFSF9 in HepG2 cells to $T{\underline{AG}}$ to create an amber stop codon. The $T{\underline{CT}}$ triplet is the codon for Ser at the $172^{nd}$ position of TNSFSF9. The two substituted nucleotides, AG, were confirmed by DNA sequencing of the PCR product followed by PCR amplification of the genomic TNFSF9 gene. Interestingly, sequencing of the cDNA of transcripts of the edited TNFSF9 gene revealed that the $T{\underline{AG}}$ had been re-edited to the wild type triplet $T{\underline{CT}}$, and 1 or 2 bases just before the triplet had been deleted. These observations indicate that CRISPR-Cas9-mediated editing of bases in target genomic DNA can be followed by spontaneous re-editing (correcting) of the bases during transcription.

Phylogenetic analysis of pleurotus species based on the nuclear SSU rRNA sequences

  • Jeong, Jae-Hoon;Kim, Eun-Kyoung;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.38-39
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    • 1996
  • The internal regions of nuclear small subunit rRNA from 6 plaeurotus species and 5 Pleurotus ostreatus strains were amplified by PCR and sequenced. The DNA sequences of 8 Pleurotus strains (P. ostreatus NFFA2, NFFA4501, NFFA4001, KFFA4001, KFCC11635, P florida, P. florida, P. sajor-cuju, P. pulmonarius, and P. spodoleucus) were idential, but P. cornucopiae differed from them in two bases out of 605 bases. However, p[hylogenetic analysis of the sequences by DNA-distance matrix and UPGMA methods showed that P. ostreatus NFFA2m1 and NFFA2m2, known as mutants of P. ostreatus NFFA2, belonged to anther group of Basidiomycotina, which is close to the genus Auricularia. The difference of the SSU rDNA sequences of P. cornucopiae from other Pleurotus species tested corresponds to the difference of mitochondrial plasmid type present in Pleurotus species as observed by Kim et al. (1993, Korean J. Microbiol. 31, 141-147).

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Phylogeny of the subfamily Salmoninae distributed in Korea based upon nucleotide sequences of mitochondrial ribosomal RNA genes (미토콘드리아 ribosomal RNA 유전자 염기서열분석에 의한 한국산 연어아과 어류의 유전적 계통도)

  • LEE Heui-Jung;PARK Jung-Youn;LEE Jeong-Ho;MIN Kwang-Sik;JEON Im Gi;YOO Mi-Ae;LEE Won-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.2
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    • pp.103-109
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    • 2000
  • Complete senuences of the mitochondrial rRNA Benes were determined among six salmonines in Korean Waters (Brachpmystax lenok, Onoorhpchus keta, O. masou mason, O. mason ishikawae, O. mykiss, and albino mutant of O. mykiss). The purposes of this study were to provide the basic information on levels of mtDNA polymorphism among these species for genetic characterization; discuss phylogentic relationships among three Oncorhynchus sepecies; demonstrate the utility of rRNA gene sequence data as a genetic marker for disringuishinf among Korean salmonines. PCR/direct sequencing data indicated the following consistent results; 1) 12S rRNA genes was 945 bases long in Oncorhynchus species, and 946 bases in B. lenot including one insertion. 2) Of sequence variation in mitochondrial rRNA regions, transitional substitutions were superior to transversion. 3) The significant differences were not shown in the intraspecific variation values in these gene regions. The percentage sequence divergence values were ranged from $0.066 to 0.212{\%}$. 4) The interspecific divergences were greater than the intraspecific variation. Nevertheless, ribosomal RMh genes were more conserved among species than the other mitochondrial genes, and they showed potentiality as an intergenic marker for systematics. In addition, phylogenetic trees, constructed from this data, supported that cherry salmon was closer to chum salmon than to rainbow trout, and that lenok was most distantly related species in six salmonid species.

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Studies on the Oranization and Expression of tRNA Genes in Aspergillus nidulans (V) The Molecular Structure of $tRNA^{Arg}$ in Aspergillus nidulans (Aspergillus nidulans의 tRNA유전자의 구조와 발현에 관한 연구 V Aspergillus nidulansd의 $tRNA^{Arg}$ 분자구조)

  • 이병재;강현삼
    • Korean Journal of Microbiology
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    • v.24 no.2
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    • pp.79-85
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    • 1986
  • We have determined the sequence of $tRNA^{Arg}$ of A. nidulans partially by enzymatic rapid RNA sequencing technique. The sequence was 5'GGCCGGCUGGCCCAAXUGGCAAGGXUCUGAXUACGAAXCAGGAGAUUGCACXXXXXGAGCXXUXXGUCGGUCACCA3' The cloverleaf structure was made from above data. As a result, the anticodon sequence was identified as ACG. This result was confirmed with charging test. The complete sequence was proposed by supplementing the DNA sequence to and by assigning the position of minor bases to this RNA sequence.

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Phylogenetic Analysis of Pleurotus Species Based on the Nuclear SSU rRNA Sequences (Phylogenetic Analysis of Pleurotus Species Based on the Nuclear SSU rRNA Sequences)

  • Jeong, Jae Hun;Kim, Eun Gyeong;No, Jeong Hye
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.37-37
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    • 1996
  • The internal regions of nuclear small subunit rRNA from 6 plaeurotus species and 5 Pleurotus ostreatus strains were amplified by PCR and sequenced. The DNA sequences of 8 Pleurotus strains (P. ostreatus NFFA2, NFFA4501, NFFA4001, KFFA4001, KFCC11635, P florida, P. florida, P. sajor-cuju, P. pulmonarius, and P. spodoleucus) were idential, but P. cornucopiae differed from them in two bases out of 605 bases. However, p[hylogenetic analysis of the sequences by DNA-distance matrix and UPGMA methods showed that P. ostreatus NFFA2m1 and NFFA2m2, known as mutants of P. ostreatus NFFA2, belonged to anther group of Basidiomycotina, which is close to the genus Auricularia. The difference of the SSU rDNA sequences of P. cornucopiae from other Pleurotus species tested corresponds to the difference of mitochondrial plasmid type present in Pleurotus species as observed by Kim et al. (1993, Korean J. Microbiol. 31, 141-147).ishement of silencing at the HMR/hsp82 locus can occur in G1-arrested cells. Cell cycle arrest at G1 phase was achieved by treatment of early log a cell cultures with .alpha.-factor mating pheromone, which induces G1 arrest. The result suggests that passage through S phase (and therefore DNA replication) is nor required for re-establishing silencer-mediated repression at the HMNRa/HSP82 locus. Finally, to test whether de nono protein synthesis is required for re-establishment of silencer-mediated repression, cells were pretreated with cycloheximide (500 /.mu.g/ml) 120 min. It was apparent that inhibiting protein synthesis delays, but does not prevent, re-establishment of silencer-mediated repression. Altogether, these results indicate that re-establishment of silencer-mediated repression is not dependent on the DNA replication and has no requirement for protein synthesis.

Interaction of Resveratrol and Genistein with Nucleic Acids

  • Usha, Subbiah;Johnson, Irudayam Maria;Malathi, Raghunathan
    • BMB Reports
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    • v.38 no.2
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    • pp.198-205
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    • 2005
  • Resveratrol (RES) and genistein (GEN) are the dietary natural products known to possess chemopreventive property and also the ability to repair DNA damage induced by mutagens/carcinogens. It is believed that the therapeutic activity of these compounds could be primarily due to their interaction with nucleic acids but detailed reports are not available. We here explore the interaction of these drugs with nucleic acids considering DNA and RNA as a potential therapeutic target. The interaction of RES and GEN has been analysed in buffered solution with DNA [saline sodium citrate (SSC)] and RNA [tris ethylene diammine tetra acetic acid (TE)] using UV-absorption and Fourier transform infrared (FTIR) spectroscopy. The UV analysis revealed lesser binding affinity with nucleic acids at lower concentration of RES (P/D = 5.00 and 10.00), while at higher drug concentration (P/D = 0.75, 1.00 and 2.50) hyperchromic effect with shift in the ${\lambda}_{max}$ is noted for DNA and RNA. A major RES-nucleic acids complexes was observed through base pairs and phosphate backbone groups with K = $35.782\;M^{-1}$ and K = $34.25\;M^{-1}$ for DNA-RES and RNA-RES complexes respectively. At various concentrations of GEN (P/D = 0.25, 0.50, 0.75, 1.00 and 2.50) hyperchromicity with shift in the ${\lambda}_{max}$ from 260 $\rightarrow$ 263 om and 260 $\rightarrow$ 270 nm is observed for DNA-GEN and RNA-GEN complexes respectively. The binding constant (from UV analysis) for GEN-nucleic acids complexes could not be obtained due to GEN absorbance overlap with that of nucleic acids at 260 nm. Nevertheless a detailed analysis with regard to the interaction of these drugs (RES/GEN) with DNA and RNA could feasibly be understood by FTIR spectroscopy. The NH band of free DNA and RNA which appeared at $3550-3100\;cm^{-1}$ and $3650-2700\;cm^{-1}$ shifted to $3450-2950\;cm^{-1}$ and $3550-3000\;cm^{-1}$ in DNA-RES and RNA-RES complexes respectively. Similarly shifts corresponding to $3650-3100\;cm^{-1}$ and $3420-3000\;cm^{-1}$ have been observed in DNA-GEN and RNA-GEN complexes respectively. The observed reduction in NH band of free nucleic acids upon complexation of these drugs is an indication of the involvement of the hydroxyl (OH) and imino (NH) group during the interaction of the drugs and nucleic acids (DNA/RNA) through H-bonded formation. The interaction of RES and GEN with bases appears in the order of G $\geq$ T > C > A and A > C $\geq$ T > G. Further interaction of these natural compounds with DNA and RNA is also supported by changes in the vibrational frequency (shift/intensity) in symmetrical and asymmetrical stretching of aromatic rings of drugs in the complex spectra. No appreciable shift is observed in the DNA and RNA marker bands, indicating that the B-DNA form and A-family conformation of RNA are not altered during their interaction with RES and GEN.

Site-Specific Mutagenesis on the 32-T and 39-T of E. coli $tRNA^{phe}$ Gene (E. coli $tRNA^{phe}$ 유전자의 32-T와 39-T 염기의 부의 특이적 돌연변이)

  • 김익영;이세영
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.176-180
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    • 1989
  • There are three pseudourdine ($Psi$)bases in the E. coli $tRNA^{phe}$ In order to study the function of the pseudouridine bases in the $tRNA^{phe}$, changes of bases $tRNA^{phe}$ gene to other bases were undertaken by the site-specific mutagenesis. Site-specific mutagenesis of T in the pheW gene, a $tRNA^{phe}$ gene of E. coli, corresponding to the baseat the No.32 position to C and also T corresponding to the base at the No.39 position to C were performed using Kunkel's uracil-containing template method. Identification of mutants were undertaken by the KNA sequencing techniques of the mutated pheW genes and activities of the mutated pheW genes complementing to E. coli NP37 mutant($pheS^{-ts}$) using the recombinant plasmid containing the mutated genes. Neither NP37 harboring pheW gene mutated at No.32 position nor NP37 harboring pheW gene mutated at No.39 position can be grown at non-permissive temperature. The result means that both mutated pheW genes can not complement to E. coli NP37, and that the pseudouridine bases are essential to the activity of the E. coli $tRNA^{phe}$ in vivo.

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A Double Helix DNA Structure Based on Block Circulant Matrix (II) (블록순환 행렬에 의한 이중나선 DNA 구조 (II))

  • Park, Ju-Yong;Kim, Jeong-Su;Lee, Moon-Ho
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.16 no.5
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    • pp.229-233
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    • 2016
  • In this paper, we present the four genetic nitrogenous bases of C, U(T), A, G to matrices and describe the structures from $4{\times}4$ RNA(ribose nucleic acid) to $8{\times}8$ DNA((deoxyribose nucleic acid) matrices. we analysis a deoxyribose nucleic acid (DNA) double helix based on the block circulant Hadamard-Jacket matrix (BCHJM). The orthogonal BCHJM is anti-symmetric pair complementary of the core DNA. The block circulant ribonucleic acid (RNA) repair damage reliability is better than the conventional double helix. In case of k=4 and N=1, the reliability of block circulant complementarity is 93.75%, and in case of k=4 and N=4, it is 98.44%. Therefore it improves 4.69% than conventional case of double helix.