• 제목/요약/키워드: cDNA sequences

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Analysis and Identification of Expressed Sequence Tags in Hairy Root Induced from Korean Ginseng (Panax ginseng C. A. Meyer)

  • Yang, Deok-Chun;In, Jun-Gyo
    • 한국약용작물학회지
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    • 제12권2호
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    • pp.154-162
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    • 2004
  • Hairy roots were induced from Korean ginseng (Panax ginseng C. A. Meyer) root explants and studied for their gene expression. A total of 3,000 ESTs (expressed sequence tags) from ginseng hairy root were determined and about 2,700 ESTs have a length of readable sequence, which result in 1,352 unique ESTs sequences. The 879 ESTs showed significant similarities to known nucleotide or amino acid sequences in other plant species, which were divided into eleven categories depending upon gene function. The remaining 473 sequences showed no significant matches, which are likely to be transcripts or to be matched to other organisms. The results indicated that the analysis of the ginseng hairy root ESTs by partial sequencing of random cDNA clones may be an efficient approach to isolate genes that are functional in ginseng root in a large scale. Our extensive EST analysis of genes expressed in ginseng hairy root not only contributes to the understanding of the dynamics of genome expression patterns in root organ but also adds data to the repertoire of all genomic genes.

Base Specificity for DNA Interstrand Cross-Linking Induced by Anticancer Agent Bizelesin

  • Lee, Chong-Soon;Myung, Pyung-Keun;Gibson, Neil W.
    • Archives of Pharmacal Research
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    • 제19권3호
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    • pp.191-196
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    • 1996
  • Bizelesin is a promising novel anticancer agent which is known to alkylate N3 of adenine to induce DNA interstrand cross-links (ISC) with in $5^I-TAATTA\; and\; 5^I-TAAAAAA$. We have investigated the base specificity for DNA ISC induced by bizelesin using oligomers containing the cross-linkable sequence $5^I-TAATTA\; and\; 5^I-TAAAAAA$. in which "N" was either A, C, G, or T. An analysis of denaturing polyacrylamide gel showed that bizelesin is able to induce DNA ISC in the duplex oligomer containing sequences $5^I-TAATTA\; and\; 5^I-TAAAAAA$. The formation of interstrand crosslinking did not occur in the sequences $5^I-TAATTA\; and\; 5^I-TAAAAAA$. DNA strand cleavage assay to determine the cross-linking site within $5^I-TAATTA$sequence showed that bizelesin alkylates guanine. These results demonstrate that bizelesin is able to induce DNA ISC at guanine but not at cytosine or thymine. In addition, guanine adducts have been found to be susceptible to DNA strand cleavage by exposure to hot piperidine. The extent of DNA strand cleavage, however, was not 100% efficient in either neutral pH buffer or hot piperidine.

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꿩에서 분리된 Newcastle Disease Virus 내열성주 (CBP)의 Fusion(F) 유전자 클론닝과 염기서열 분석 (Molecular Cloning and Nucleotide Sequence of the Gene Encoding Fusion(F) Protein of the Thermostable Newcastle Disease Virus Isolated from a Diseased Pheasant)

  • 장경수;전무형;송희종;김귀현;박종현
    • 대한바이러스학회지
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    • 제28권3호
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    • pp.233-245
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    • 1998
  • The gene encoding F protein of CBP-1 strain, a heat-stable Newcastle disease virus (NDV) isolated from the diseased pheasants in Korea, was characterized by reverse transcription-polymerase chain reaction (RT-PCR), nucleotide and amino acid sequences. Virus RNA was prepared from the chorioallatoic fluid infected with NDV CBP-1 virus and cDNA was amplified by RT-PCR, cloned and sequenced to analyze. The PCR was sensitive as to detect the virus titer above $2^5$ hemagglutination unit. 1.7kb (1,707bp) size of the cDNA was amplified and cloned into BamHI site of pVL1393 Baculo transfer vector. The nucleotide sequences for F protein were determined by dye terminator cyclic sequencing using four pairs of primers, and 553 amino acid sequences were predicted. In comparison of the nucleotide sequence of F gene of CBP-1 with those of other NDV strains, the homology revealed 88.8%, 98.5% and 98.7% with Kyojungwon (KJW), Texas GB and Beaudette C strains, respectively. As the deduced 553 amino acid sequences of F protein of CBP-1 were compared with those of other NDV strains, the homology appeared 89.9%, 98.7% and 98.9% with KJW, Texas GB and Beaudette C strains, respectively. The putative protease cleavage site (112-116) was R-R-Q-K-R, indicating that CBP-1 strain is velogenic type. The amino acid sequences include 6 sites of N-asparagine-linked glycosylation and 13 cysteine residues. These data indicate that the genotype of CBP-1 strain is more closely associated with the strains of Texas GB and Beaudette C than KJW strain.

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nrDNA-ITS 분자마커를 이용한 오미자(五味子) 종 감별 및 기원분석 -ITS 염기서열을 이용한 오미자(五味子) 감별- (Molecular Authentication of Schisandrae Fructus and Analysis of Phylogenetic Relationship based on nrDNA-ITS sequences)

  • 문병철;지윤의;서형석;이아영;천진미;김호경
    • 대한본초학회지
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    • 제25권4호
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    • pp.47-54
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    • 2010
  • Objectives : The original plant species of Schisandrae Fructus (O-mi-ja) is prescribed as Schisandra chinensis $B_{AILL.}$, in Korea, but S. chinensis $B_{AILL.}$ and S. sphenanthera $R_{EHD.}$ et $W_{ILS.}$ in China. Moreover, fruit of several other species in genus Schisandra also have been used as the same herbal medicines. To develop a reliable method for correct identification of Schisandrae Fructus and to evaluate the phylogenetic relationship of S. chinensis and its related species, we analyzed internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA (nrDNA). Methods : Twenty-four plant samples of three Schisandra species and one Kadsura species, S. chinensis $B_{AILL.}$, S. spenanthera $R_{EHD.}$ et $W_{ILS.}$, S. nigra $M_{ax.}$ and Kadsura japonica $D_{UNAL}$ were collected from each different native habitate and farm in Korea and China. The nrDNA-ITS region of each samples were amplified using ITS1 and ITS4 primer and nucleotide sequences were determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW based on the entire nrDNA-ITS sequence. Results : In comparative analysis of the nrDNA-ITS sequences, we found specific nucleotide sequences including indels (insertions and deletions) and substitutions to distinguish C. chinensis, S. spenanthera, S. nigra, and K. japonica. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origin of O-mi-ja. Moreover, we evaluated the phylogenetic relationship of four plant species by the analysis of nrDNA-ITS sequences. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by the providing of definitive information that can identify each plant species and distinguish it from unauthentic adulterants for O-mi-ja.

Cloning and Characterization of cDNA Encoding Potentially Functional Mouse Glandular Kallikrein

  • Kim, Hwa-Seon;Kim, Won-Sin
    • BMB Reports
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    • 제30권5호
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    • pp.356-361
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    • 1997
  • We cloned a cDNA (pPRC-1) which was comprised of 841 nucleotides from the cDNA library of a male ICR mouse submandibular gland ($SMG^+$). The nucleotide sequences of pPRC-1 were identical to those of exons 2 and 3 of the mGK-21 gene, a potentially functional glandular kallikrein identified in a Balb/c mouse, except for one nucleotide residue. Although this substitution changes Ile (ATT) in pPRC-1 to Val (GTT) in mGK-21, this difference has been explained by strain polymorphism. From the amino acid sequences predicted from its cDNA, we speculated that mGK-21 gene products/pGK21 consist of 261 amino acids including the $NH_2$-terminal signal peptide (residues 1~17), the short propeptide (residues 17~24), and the active peptide (residues 25~261). Although we did not demonstrate the enzyme activity of pGK21, it was assumed that pGK 21 was involved in the maturation of certain bioactive polypeptide(s) in mouse SMG for the following reasons : (a) mGK-21 gene was apparently expressed in a male ICR mouse SMG: (b) the proposed active site $His^{65}$, $Asp^{120}$, and $Ser^{213}$ residues were completely conserved in pGK21 just like other glandular kallikreins; (c) the cloned cDNA was translated to a predicted 27 kDa polypeptide chain in vitro: (d) the 27 kDa polypeptide chain produced by CHO cells was produced to a putative active form by trypsin.

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엽록체 DNA trnL-F 및 핵리보조옴 DNA ITS, ETS 염기서열에 의한 고려인삼의 유전적 변이와 계통학적 유연관계 (Genetic Variation and Phylogenetic Relationship of Korean Ginseng based on cpDNA trnL-F, nrDNA ITS and ETS Sequences)

  • 방찬국;김주환;백명현;김창식;엄동명;김동희
    • 동의생리병리학회지
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    • 제18권6호
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    • pp.1699-1709
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    • 2004
  • Genetic variations and relationship based on the sequences of cpDNA trnL-F gene, nrDNA ITS and ETS region among the twenty four taxa including Panax ginseng C.A. Meyer and its related species were investigated. And taxonomic status and molecular phylogenetic relationship between P. ginseng and related groups were discussed. Molecular systematic data from cpDNA and nrDNA sequences were very useful to elucidate the genetic variations and relationships among the treated taxa. It was found that P. ginseng is the independent unique species with distinct genetic limitation from the related species such as P. quinquefolius, P. japonicum, P. notoginseng and P. pseudoginseng. P. ginseng including cultivated types as well as wild ones formed monophyletic group with high genetic similarities. P. quinquefolius and P. japonicum were the most related sister groups of P. ginseng based on the molecular phylogenetic results in this study.

Identification of Luteovirus Nucleotide Sequences in Mild Yellow-Edge Diseased Strawberry Plants

  • Shaban Montasser, Magdy;Al-Awadhi, Husain;Hadidi, Ahmed
    • The Plant Pathology Journal
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    • 제18권1호
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    • pp.1-5
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    • 2002
  • The availability of nucleotide sequences of the coat protein gene of Potato leafroll virus (PLRV) permitted the construction of DNA primers that were utilized for cDNA synthesis. Polymerase chain reaction (PCR) products of a 487 bp. and approximately 500 bp DNA fragments were amplified from nucleic acid extracts of PLRV-infected tissue and strawberry mild yellow-edge (SMYE) diseased strawberry tissue, respectively. The amplified DNA fragments were further differentiated by hybridization analysis with a CDNA probe for the coat protein gene of PLRV and restriction fragment length polymorphism (RFLP) analysis. These results suggest that a luteovirus is associated with the SMYE disease.

Molecular Phylogeny of Silk Producing Insects Based on Internal Transcribed Spacer DNA1

  • Mahendran, Botlagunta;Ghosh, Sudip K.;Kundu, Subhas C.
    • BMB Reports
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    • 제39권5호
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    • pp.522-529
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    • 2006
  • Silk moths are the best studied silk secreting insects and belong to the families Bombycidae and Saturniidae. The phylogenetic relationship between eleven silk producing insects was analyzed using the complete DNA sequence of the internal transcribed spacer DNA 1 locus. The PCR amplification and sequence analysis showed variation in length ranging from 138 bp (Antheraea polyphemus) to 911 bp (Hyalopora cecropia). Microsatellite sequences were found and was be used to distinguish Saturniidae and Bombycidae members. The nucleotide sequences were aligned manually and used for construction of phylogenetic trees based on Maximum parsimony and Maximum likelihood methods. The topology in both the approaches yielded a similar tree that supports the ancestral position of the Antheraea assama.

Re-description of Chrysaora pacifica (Goette, 1886) (Cnidaria, Scyphozoa) from Korean Coastal Waters: Morphology and Molecular Comparisons

  • Lee, Hye Eun;Yoon, Won Duk;Chae, Jinho;Ki, Jang-Seu
    • Ocean and Polar Research
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    • 제38권4호
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    • pp.295-301
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    • 2016
  • The nomenclature of the sea nettle jellyfish from Korea was initially described as Dactylometra quinquecirrha Agassiz, 1862. However its identity has been questioned on the basis of its local distribution and molecular data. Here, we examined morphology and DNA sequences of nuclear rDNA using specimens collected from southern Korean waters in August 2014. Based on morphological characteristics (bell size, umbrella pattern, number of tentacles and lappets) and distribution locality, we reassign the Korean D. quinquecirrha to Chrysaora pacifica (Goette, 1886), and provide a re-description accordingly. The molecular identity of C. pacifica was further confirmed by comparison of nuclear ribosomal DNA sequences.