• 제목/요약/키워드: Nucleotides

검색결과 848건 처리시간 0.023초

멧누에(Bombyx mandarina)로부터 $\beta$-N-Acetyglunosamicidase를 코딩하는 cDNA의 분리 및 염기서열 결정 (Molecular Cloning and Characterization of the Gene encoding $\beta$-N-acetylhlucosaminidases Homologue from Bombyx mandarina)

  • 구태원;황재삼;성규병;윤은영;방혜선;권오유
    • 한국잠사곤충학회지
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    • 제41권3호
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    • pp.147-153
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    • 1999
  • Chitinolytic enzymes such as ${\beta}$-N-acetylglucosaminidase are major hydrolases involved in insect molting. We have isolated, sequenced a CDNA encoding ${\beta}$-N-acetylglucosaminidase from the silkworm, Bombyx mandarina, and compared its sequence with genes encoding chitinolytic enyzmes from other sources. The insert DNA in the clone is 3,284 nucleotides long with an open reading frame of 1,788 nucleotides that encodes a protein of 596 amino acids with a molecular weight of 68.2 kDa. There is a 3’-untranslated region composed with 1.479 nucleotides and are several potential polyadenylation signals. The predicted amino acid sequence apparently contains a leader peptide of 23 amino acids. A search of the amino acids sequence databases for sequences similarities to other ${\beta}$-N-acetylglucosaminidases or ${\beta}$-N-acetylhexosaminidases. The highest similarity matched with the enzyme from B. mori, which has a sequence identity of 95%. On the other hand, the identity between the B. mandarina enzyme and those from M. sexta and human are 70% and 24%, respectively.

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Full-Length cDNA Cloning and Nucleotide Sequence Analysis of Cucumber Mosaic Virus (Strain Kor) RNA2

  • Kwon, Chang-Seob;Park, Kyung-Hee;Chung, Won-Il
    • Journal of Plant Biology
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    • 제39권4호
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    • pp.265-271
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    • 1996
  • Full-length cDNA for RNA2 of cucumber mosaic virus strian Kor (Kor-CMV) was cloned downstream of synthetic T7 promoter by reverse transcriptase-polymerase chain reaction (RT-PCR). The clone could generate a full-length transcript corresponding to RNA1 in size when synthesized by T7 RNA polymerase. The complete nucleotide sequence has shown that the RNA2 is composed of 3,049 nucleotides and contains one functional open reading frame (ORF) of 2,574 nucleotides encoding 2a protein. The deduced translation product of the 2,574 nucleotides contains GDD motif which is a characteristic of RNA-dependent RNA polymerase (RdRp). The amino acid sequence analysis of the 2a protein has shown that the homology is found in decreasing order with O-CMV (98.8%), Y-CMV (98.7%), Fny-CMV (98.3%), KCMV (94.9%), Ix-CMV (91.9%), and Q-CMV (74.9%). Kor-CMV is suggested to belong to subgroup Ⅰ in the aspect of nucleotide sequence homology of RNA2.

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DNA 바코드 분석을 통한 패장 기원종 감별용 분자 마커 개발 (Development of Molecular Marker for the authentication of Patriniae Radix by the analysis of DNA barcodes)

  • 김욱진;지윤의;이영미;강영민;최고야;김호경;문병철
    • 대한본초학회지
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    • 제29권6호
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    • pp.45-53
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    • 2014
  • Objectives : Due to the morphological similarity of in the roots of herbal medicine, the official herbal medicine is very difficult to authenticate between the original plants of Patriniae Radix and two adulterant Patrinia species. Therefore, we introduced DNA barcode analysis to establish a powerful tool for the authentication of Patriniae Radix from its adulterants. Methods : To analyze DNA barcode regions, genomic DNA was extracted from twenty-nine specimens of Patrinia scabiosaefolia, Patrinia villosa, Patrinia saniculifolia, and Patrinia rupestris, and internal transcribed spacer 2(ITS2), matK and rbcL genes were amplified. For identification of species specific sequences, a comparative analysis was performed by the ClastalW based on entire sequences of ITS2, matK and rbcL genes, respectively. Results : In comparison of three DNA barcode sequences, we identified 22, 22, and 12 species-specific nucleotides enough to distinguish each four species from ITS2, matK and rbcL gene, respectively. The sequence differences at the corresponding positions were available genetic marker nucleotides to discriminate the correct species among analyzed four species. These results indicated that comparative analysis of ITS2, matK and rbcL genes were useful genetic markers to authenticate Patriniae Radix. Conclusions : The marker nucleotides enough to distinguish P. scabiosaefolia, P. villosa, P. saniculifolia, and P. rupestris, were obtained at 22 SNP marker nucleotides from ITS2 and matK DNA barcode sequences, but they were confirmed at 12 SNP marker nucleotides from rbcL. These differences could be used to authenticate Patriniae Radix from its adulterants as well as discriminating each four species.

Effect of Glutamine, Glutamic Acid and Nucleotides on the Turnover of Carbon (δ13C) in Organs of Weaned Piglets

  • Amorim, Alessandro Borges;Berto, Dirlei Antonio;Saleh, Mayra Anton Dib;Telles, Filipe Garcia;Denadai, Juliana Celia;Sartori, Maria Marcia Pereira;Luiggi, Fabiana Golin;Santos, Luan Sousa;Ducatti, Carlos
    • Asian-Australasian Journal of Animal Sciences
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    • 제29권8호
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    • pp.1152-1158
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    • 2016
  • Morphological and physiological alterations occur in the digestive system of weanling piglets, compromising the performance in subsequent phases. This experiment aimed at verifying the influence of glutamine, glutamate and nucleotides on the carbon turnover in the pancreas and liver of piglets weaned at 21 days of age. Four diets were evaluated: glutamine, glutamic acid or nucleotides-free diet (CD); containing 1% glutamine (GD); containing 1% glutamic acid (GAD) and containing 1% nucleotides (ND). One hundred and twenty-three piglets were utilized with three pigs slaughtered at day zero (weaning day) and three at each one of the experimental days (1, 2, 4, 5, 7, 9, 13, 20, 27, and 49 post-weaning), in order to collect organ samples, which were analyzed for the ${\delta}^{13}C$ isotopic composition and compared by means of time. No differences were found (p>0.05) among treatments for the turnover of the $^{13}C$ in the pancreas ($T_{50%}$ = 13.91, 14.37, 11.07, and 9.34 days; $T_{95%}$ = 46.22, 47.73, 36.79, and 31.04 days for CD, GD, GAD, and ND, respectively). In the liver, the ND presented accelerated values of carbon turnover ($T_{50%}=7.36$ and $T_{95%}=24.47days$) in relation to the values obtained for the GD ($T_{50%}=10.15$ and $T_{95%}=33.74days$). However, the values obtained for the CD ($T_{50%}=9.12$ and $T_{95%}=30.31days$) and GAD ($T_{50%}=7.83$ and $T_{95%}=26.03days$) had no differences (p>0.05) among other diets. The technique of $^{13}C$ isotopic dilution demonstrated trophic action of nucleotides in the liver.

Sequencing of cDNA Clones Expressed in Adipose Tissues of Korean Cattle

  • Bong, J.J.;Tong, K.;Cho, K.K.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • 제18권4호
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    • pp.483-489
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    • 2005
  • To understand the molecular mechanisms that regulate intramuscular fat deposition and its release, cDNA clones expressed in adipose tissues of Korean cattle were identified by differential screening from adipose tissue cDNA library. By partial nucleotide sequencing of 486 clones and a search for sequence similarity in NCBI nucleotide databases, 245 clones revealed unique clones. By a functional grouping of the clones, 14% of the clones were categorized to metabolism and enzyme-related group (stearoyl CoA desaturase, lactate dehydrogenase, fatty acid synthase, ATP citrate lyase, lipoprotein lipase, acetyl CoA synthetase, etc), and 6% to signal transduction/cell cycle-related group (C/EBP, cAMP-regulated phosphoprotein, calmodulin, cyclin G1, cyclin H, etc), and 4% to cytoskeleton and extracellular matrix components (vimentin, ankyrin 2, gelosin, syntenin, talin, prefoldin 5). The obtained 245 clones will be useful to study lipid metabolism and signal transduction pathway in adipose tissues and to study obesity in human. Some clones were subjected to full-sequencing containing open reading frame. The cDNA clone of bovine homolog of human prefoldin 5 gene had a total length of 959 nucleotides coding for 139 amino acids. Comparison of the deduced amino acid sequences of bovine prefoldin 5 with those of human and mouse showed over 95% identity. The cDNA clone of bovine homolog of human ubiquitin-like/S30 ribosomal fusion protein gene had a total length of 484 nucleotides coding for 133 amino acids. Comparison of the deduced amino acid sequences of bovine ubiquitin-like/S30 ribosomal fusion protein gene with those of human, rat and mouse showed over 97% identity. The cDNA clone of bovine homolog of human proteolipid protein 2 mRNA had a total length of 928 nucleotides coding for 152 amino acids. Comparison of the deduced amino acid sequences of bovine proteolipid protein 2 with those of human and mouse showed 87.5% similarity. The cDNA clone of bovine homolog of rat thymosin beta 4 had a total length of 602 nucleotides coding for 44 amino acids. Comparison of the deduced amino acid sequences of bovine thymosin beta 4 gene with those of human, mouse and rat showed 93.1% similarity. The cDNA clone of bovine homolog of human myotrophin mRNA had a total length of 790 nucleotides coding for 118 amino acids. Comparison of the deduced amino acid sequences of bovine myotrophin gene with those of human, mouse and rat showed 83.9% similarity. The functional role of these clones in adipose tissues needs to be established.

Allosteric Properties of Hafnia alvei Aspartase by Nucleotide Effectors

  • Noh, Hak-Joon;Kwon, Si-Joong;Kim, Ki-Tae;Lee, Chang-Hyun;Yoon, Moon-Young
    • BMB Reports
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    • 제33권5호
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    • pp.366-369
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    • 2000
  • The nucleotide effects of Hafnia alvei aspartase were investigated. Purine nucleosides, such as adenosine and guanosine, increased the aspartase activities; whereas, purine nucleotides, such as AMP, ATP, GTP and IMP, caused little change in the aspartase activities. However, pyrimidine derivatives, such as cytidine and CTP, decreased the aspartase activity. The nucleotide and nucleoside effects by the limited trypsin-treated aspartase were similar to those of a native enzyme. These results indicate that the COOH-terminal region and an allosteric site might be located away from each other. The initial velocity study in the presence of adenosine showed that $K_m$ for aspartate was decreased to one-sixth of that in the absence of adenosine, but $V_{max}$ was unchanged. The significance of the distinct allosteric effect for the enzyme-nucleotide interaction is discussed.

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

  • 구태원;황재삼;성규병;윤은영;방혜선;권오유
    • 생명과학회지
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    • 제9권4호
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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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Characterization of a Noncanonical Purine dNTP Pyrophosphatase from Archaeoglobus fulgidus

  • Im Eun-Kyoung;Hong Chang-Hyung;Back Jung-Ho;Han Ye-Sun;Chung Ji-Hyung
    • Journal of Microbiology and Biotechnology
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    • 제16권7호
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    • pp.1144-1148
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    • 2006
  • DNA can oxidatively be deaminated by ROS, which converts DNA base amino groups to keto groups and can trigger abnormal mutations, resulting in mutagenesis in organisms. In this study, a noncanonical purine dNTP pyrophosphatase (AfPPase) from a hyperthermophilic archaeon Archaeoglobus fulgidus, which hydrolyzes aberrant nucleoside triphosphates, was overexpressed in E. coli, purified, and characterized. The purified AfPPase showed remarkably high activity for XTP and dITP, suggesting that the 6-keto group of these nucleotides is critical for the reactivity. Under optimal reaction conditions, the reaction rate for these substrates was about 120 times that with dGTP. Therefore, AfPPase may play a significant role in DNA repair by hydrolysis of noncanonical nucleotides before they are misincorporated into DNA.