• Title/Summary/Keyword: Adenine Nucleotide

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Comparative Analysis of Nucleotide Sequence and Codon Usage of Arylphorin Gene Cloned from Four Silk-Producing Insects and Their Molicular Phylogenetics

  • Lee, Sang-Mong;Hwang, Jae-Sam;Lee, Jin-Sung;Goo, Tae-Won;Kwon, O-Yu;Kim, Ho-Rak
    • Journal of Life Science
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    • v.9 no.1
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    • pp.84-89
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    • 1999
  • To determine phylogenetic relatedness of four silk-producing silkmoths (B. mori, B. mandarina, A. yamamai and A. pernyi), internal coding region of arylphorin which is a storage protein in hemolymph protein of insects were amplified by polymerase chain reaction and then sequenced and compared each other. The nucleotide composition was biased toward adenine and thymine(59% A+T) and a strong bias for use of C in the third position of codons was found for Phe and Tyr. Together TTC(Phe) and TAC(Tyr) account for about 16.8% (10 for TTC and 8 for TAC) of all codon usage. The nucleotide similarity of arylphorin gene from B. mori showed 99%, 98% and 97% homology with those of B. mandarina, A. yamamai and A. pernyi, respectively. Also, the nucleotide sequence of arylphorin gene from B. mandarina showed 98% and 97% homology with those of A. yamamai and A.pernyi, respectively. Between A. yamamai and A. pernyi, the sequence homology was 97%. The deduced amino acid sequences in B. mori, B. mandarina and A. yamamai showed almost 99% homology. Although the aryphorin gene provided insufficient variability among the four insect species, A UPGMA tree is generated that supported the monophyly of silk-producing insects, with M. sexta placed basal to it. It is suggest that silk-producing insects have a close relationship and a homogeneous genetic background from comparison with those of other insects.

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19F NMR Investigation of F1-ATPase of Escherichia coli Using Fluorinated Ligands

  • Jung, Seun-Ho;Kim, Hyun-Won
    • BMB Reports
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    • v.31 no.1
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    • pp.44-47
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    • 1998
  • Asymmetry amongst nucleotide binding sites of Escherichia coli $F_1$-ATPase was examined using $^{19}F$ NMR signal from fluorinated analogs of adenine nucleotides bound to nucleotide binding sites. ADP-$CF_2-{PO_3}^{2-}$ showed no inhibitory effect to $F_1$-ATPase. But ADP-CHF-${PO_3}^{2-}$ (racemic mixture) showed competitive inhibition of $F_1$-ATPase with $K_i$ of $60\;{\mu}m$. ADP-CHF-${PO_3}^{2-}$ shows only negligible binding to $EF_1$ in the absence of $Mg^2+$. With the addition of $Mg^2+$ to the medium, the $^{19}F$ resonance of free ADP-CHF-${PO_3}^{2-}$ disappeared and the new broad resonances appeared. Appearance of more than two new asymmetric resonances following the binding of ADP-CHF-${PO_3}^{2-}$ to $EF_1$ may indicate that at least one of the isomers showed split resonances. This may suggest that the region between ${\alpha}$-and ${\beta}$-phosphate of ADP-CHF-${PO_3}^{2-}$ which is bound to catalytic sites is experiencing a different environment at different sites.

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Analysis for nucleotide sequence of the small membrane (sM) protein gene of porcine epidemic diarrhea virus Chinju99 isolated in Korea

  • Yeo, Sang-Geon;Lee, Changhee
    • Korean Journal of Veterinary Research
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    • v.47 no.2
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    • pp.169-174
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    • 2007
  • To provide information for the molecular pathogenesis and antigenic structures of Korean isolates of porcine epidemic diarrhea virus (PEDV), the small membrane (sM) protein gene of Chinju99 strain, which was previously isolated from piglets suffering from severe diarrhea was characterized and further analyzed with other PEDV strains. The sM gene of Chinju99 generated by reverse transcription and polymerase chain reaction had a single open reading frame with 231 bases consisting of 24.2% adenine, 18.6% cytosine, 18.1% guanine and 39.0% thymine nucleotides. Nucleotide sequence of the gene revealed 97.8% homology to those of Belgian strain CV777 and British strain Br1/87, and 97.0% to Chinese strain LZC. The gene encoded a protein with 76 amino acids, and putative amino acid sequence of the gene revealed 98.7% homology to those of CV777 and Br1/87, and 96.1% to LZC. The amino acids of Chinju99 sM gene consisted of mostly hydrophobic residues, and there were one potential N-myristylation site and one potential threonine (T)-linked phosphorylation site recognized. Also, there was a transmembrane region with 46 amino acids, and Chinju99 was more close to CV777 and Br1/87 than to LZC in phylogenetic analysis on the sM amino acid sequences.

A Case of Leber's Hereditary Optic Neuropathy Showing 11778 Point Mutation of Mitochondrial DNA (사렵체 DNA의 11778 점돌연변이가 확인된 Leber씨 유전성 시신경병증 1례)

  • Jung, Yun-Seok;Park, Seung-Kwon;Lee, Seung-Yeop;Hah, Jung-Sang;Park, Mee-Yeoung;Lee, Se-Jin;Lee, Jun
    • Journal of Yeungnam Medical Science
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    • v.16 no.1
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    • pp.114-118
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    • 1999
  • Leber's hereditary optic neuropathy(LHON) is an optic nerve disease that causes blindness and is associated with maternally inherited mitochondrial DNA(mt DNA) mutations. The most common mitochondrial DNA mutation among LHON patients is a point mutation at the nucleotide 11778 in the subunit 4 of complex I. In one 45-year old male LHON patient with bilateral optic neuropathy. we investigated the presence of a point mutation of mitochondrial DNA and identified a single guanine to adenine transition mutation in the mitochondrial DNA at nucleotide point 11778.

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Characterization of the Stearic Acid-Induced Uncoupling of Mitochondrial Respiration

  • Chun, Sun-Bum;Ho, Sung-Sook;Han, Sang-Hwa
    • BMB Reports
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    • v.30 no.2
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    • pp.138-143
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    • 1997
  • In order to assess controversial' proposals concerning the fatty acid-induced uncoupling of mitochondrial oxidative phosphorylation, we investigated the interaction of stearic acid with key mitochondrial proteins and measured the effect of stearic acid on the respiration of cytochrome c oxidase vesicles. Electron paramagnetic resonance spectra of spin-labeled stearic acid clearly demonstrated that cytochrome c oxidase interacts strongly with stearic acid. However, the respiration of detergent-solubilized cytochrome c oxidase was not altered significantly by stearic acid. Surprisingly, adenine nucleotide carrier, which was assumed to bind and translocate fatty acid anions in the Skulachev model of uncoupling, did not bind stearic acid at all. The respiration rate of cytochrome c oxidase vesicles was increased by ~70% in the presence of $20{\mu}m$ stearic acid and this uncoupling was attributed to a simple protonophoric effect of stearic acid.

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Purification and characterization of glutamine phosphoribosylpyrophosphate amidotransferase from streptomyces tubercidicus (Streptomyces tubercidicus에서 glutamine phosphoribosylpyrophosphate amidotransferase의 정제 및 특성)

  • 하영칠;유진철
    • Korean Journal of Microbiology
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    • v.29 no.2
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    • pp.97-103
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    • 1991
  • Glutamine phosphoribosylpyrophosphate amidotransferase of Streptomyces tubercidicus was purified and characterized. Molecular weight of the isolated enzyme was determined to be approximately 230,000 and was composed foru identical subunits having a molecular weight of 58,000. This enzyme was strongly inhibited by AMP while considerably inhibited by ATP and GTP. Inhibition effect of enzyme activity by AMP was antagonized by increased concentration of substrate, PRPP, and metal ion (especially, $Mg^{++}$) was essential in both catalytic activity and nucleotide inhibition of this enzyme. Therefore, it was confirmed that end product inhibition of glutamine phosphoribosylpyrophosphate amidotransferase by adenine participated in the regulation of tubercidin biosynthesis from Streptomyces tubercidicus.s.

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Identification of a Tandemly Repented DNA Sequence Using Combined RAPD and FISH in Welsh Onion (Allium fistulosum)

  • Bong Bo Seo;Geum Sook Do;Seon Hee Lee
    • Animal cells and systems
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    • v.3 no.1
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    • pp.69-72
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    • 1999
  • A tandemly repeated DNA sequence was identified and characterized y the combined RAPD and FISH data from a total genomic DNA of Welsh onion (Allium fistulosum). A clone containing this repeating sequence was selected and sequenced. This repeating unit of 314 bp inserted into pAf 072 contained 54.1% adenine and thymine residues, and showed the primer sequence used, 5'-GAAACGGGTG-3', in both terminals of the sequence. Fluorescence in situ hybridization using this tandemly repeated sequence as a probe indicated that the detected sites were coincident with the major C-banded constitutive heterochromatin in the terminal regions of both arms of all 6 chromosomes.

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Characterization of 18S rDNA in Polygonatum spp. Collections (둥굴레속 식물의 18S rDNA 염기서열의 특성)

  • Yun, Jong-Sun;Kim, Ik-Hwan;Park, Jae-Seong;Lee, Cheol-Hee;Hong, Eui-Yon;Yun, Tae;Jong, Seung-Keun
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.3
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    • pp.178-182
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    • 2006
  • This study was conducted to investigate the variation in sequence, the base composition and the sequence similarity of 18S rDNA (18S ribosomal RNA coding region) in the 10 Polygonatum spp. collections. The entire 18S rDNA region of 10 Polygonatum spp. collections ranged from 913 bp to 914 bp. There were 8 variable sites in the entire 18S region, and they were attributable to nucleotide substitution and deletion. $T{\rightarrow}C$ transition happened in 4 sites, and $A{\rightarrow}G$ transition happened in 1 site. $C{\rightarrow}A$ transversion happened in 1 site, and deletion happened in 2 sites. Transition rates were five times that of transversion. Base compositions of 18S rDNA were $23.09{\sim}23.33%$ in adenine, $23.33{\sim}23.52%$ in guanine, $25.60{\sim}25.85%$ in thymine and $27.38{\sim}27.79%$ in cytosine. The A + T content of 18S rDNA of 10 Polygonatum spp. collections averages 48.99%, ranging from 48.80% to 49.18%, and the G + C content averages 51.01%, ranging from 50.82% to 51.20%. Pairwise sequence comparisons indicated that 18S rDNA sequence similarity ranged from 99.7% to 100%.

Expressed Sequence Tag Analysis for Identification and Characterization of Sex-Related Genes in the Giant Tiger Shrimp Penaeus monodon

  • Preechaphol, Rachanimuk;Leelatanawit, Rungnapa;Sittikankeaw, Kanchana;Klinbunga, Sirawut;Khamnamtong, Bavornlak;Puanglarp, Narongsak;Menasveta, Piamsak
    • BMB Reports
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    • v.40 no.4
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    • pp.501-510
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    • 2007
  • Sex-related genes expressed in vitellogenic ovaries of the giant tiger shrimp, Penaeus monodon, were identified by an EST approach. A total of 1051 clones were unidirectionally sequenced from the 5 terminus. Nucleotide sequences of 743 EST (70.7%) significantly matched known genes previously deposited in the GenBank (E-value <$10^{-4}$) whereas 308 ESTs (29.3%) were regarded as newly unidentified transcripts (E-value >$10^{-4}$). A total of 559 transcripts (87 contigs and 472 singletons) were obtained. Thrombospondin (TSP) and peritrophin (79 and 87 clones accounting for 7.5 and 8.3% of clones sequenced, respectively) predominated among characterized transcripts. everal full length transcripts (e.g. cyclophilin, profillin and thioredoxin peroxidase) were also isolated. A gene homologue encoding chromobox protein (PMCBX, ORF of 567 nucleotides encoding a protein of 188 amino acids) which is recognized as a new member of the HP1 family was identified. Expression patterns of 14 of 25 sex-related gene homologues in ovaries and testes of P. monodon broodstock were examined by RT-PCR. Female sterile and ovarian lipoprotein receptor homologues were only expressed in ovaries whereas the remaining transcripts except disulfide isomerase related P5 precursor and adenine nucleotide translocator 2 were higher expressed in ovaries than testes of P. monodon broodstock. A homologue of ubiquitin specific proteinase 9, X chromosome (Usp9X) revealed a preferential expression level in ovaries than testes of broodstock-sized P. monodon (N = 13 and 11, P<0.05) but was only expressed in ovaries of 4-month-old shrimp (N = 5 for each sex).

Sequence Analysis, Molecular Cloning and Restriction Mapping of Mitochondreal Genome of Domesticated Silkworm, Bombyx mori (누에 미토콘드리아 유전체의 제한효소 지도작성, 클로닝 및 염기서열 분석)

  • 이진성;성승현;김용성;서동상
    • Journal of Sericultural and Entomological Science
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    • v.42 no.1
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    • pp.14-23
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    • 2000
  • The mitochondrial genome of domesticated silkworm (Bombyx mori) was mapped with five restriction endonucleases (BamHI, EcoRI, HindIII, PstI and XbaI), the entire genome was cloned with HindIII and EcoRI. From the end sequencing results of 5$^1$and 3$^1$region for full genome set of eleven mitochondrial clones, the seven mitochondrial genes (NADH dehydrogenase 6, ATPase 6, ATPase 8, tRN $A^{Lys}$, tRN $A^{Asp}$, tRN $A^{Thr}$ and tRN $A^{Phe}$ of mori were identified on the basis of their nucleotide sequence homology. The nucleotide composition of NADH dehydrogenase 6 was heavily biased towards adenine and thymine, which accounted for 87.76%. On basis of the sequence similarity with published tRNA genes from six insect species, the tRN $A^{Lys}$, tRN $A^{Asp}$ and tRN $A^{Thr}$ were showed stable canonical clover-leaf tRNA structures with acceptible anticodons. However, both the DHU and T$\psi$C arms of tRN $A^{Phe}$ could not form any stable stem-loop structure. The two overlapping gene pairs (tRN $A^{Lys}$ -tRN $A^{ASP}$ and ATPase8-ATPase6) were found from our sequencing results. The genes are encoded on the same strad. ATPase8 and ATPase6 overlaps (ATGATAA) which are a single example of overlapping events between abutted protein-coding genes are common, and there is evidence that the two proteins are transcribed from a single bicistronic message by initiation at 5$^1$terminal start site for ATPase8 and at an internal start site for ATPase6. Ultimately, this result will provide assistance in designing oligo-nucleotides for PCR amplification, and sequencing the specific mitochondrial genes for phylogenetics of geographic races, genetically improved silkworm strains and wild silkworm (mandarina) which is estimated as ancestal of domesticated silkworm.sticated silkworm.

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