• Title/Summary/Keyword: Cloning of R Genes

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Molecular Cloning and Characterization of a Heat Shock Protein 70-related cDNA from Olive flounder (Paralichthys olivaceus)

  • Kim, Woo-Jin;Lee, Jeong-Ho;Kim, Kyung-Kil;Lee, Sang-Jun;Kang, Ho-Sung;Kim, Han-Do
    • Journal of Aquaculture
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    • v.12 no.2
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    • pp.91-100
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    • 1999
  • The complete nucleotide sequence of olive flounder (Paralichthys olivaceus) hsp70-related rDNA was determined by RT- and RACE-PCR methods. A full-length of hsp70-related cDNA has an open reading frame of 1.95 kb encoding 650 amino acids with a calculated molecular weight of 71.1 kD. A corresponding hsp70-related protein contains a number of conserved elements including an ATP-binding domain, a nuclear localization signal and the carboxyl terminal motif, EEVD, which may have a role in chaperone function. Comparison of nucleotide and predicted amino acid sequence between olive flounder hsp70-related gene and hsp/hsc70 genes of other species revealed a high similarity with the cognate form of these genes. These results indicated that we recovered likely to be a olive flounder cognate hsc70 gene.

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Cloning and Characterization of Genes Controlling Flower Color in Pharbitis nil Using AFLP (Amplified Fragment Length Polymorphism) and DDRT (Differential Display Reverse Transcription)

  • Kim, Eun-Mi;Jueson Maeng;Lim, Yong-Pyo;Yoonkang Hur
    • Journal of Photoscience
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    • v.7 no.2
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    • pp.73-78
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    • 2000
  • To analyze molecular traits determining pigmentation between Pharbitis nill violet and white, Amplified Fragment Length Polymorphism(AFLP) and Differential Display Reverse Transcription(DDRT) experiments were carried out with either genomic DNAs or total RNAs isolated from both plants. Results of AFLP experiment in combination of 8 EcoRⅠ primers with 6 MseⅠ primers showed 41 violet-and 60 white-specific DNA bands. In the subsequent experiment, 22 violet-and 22 white-specific DNA fragments were amplified by PCR with DNAs eluted. The sizes of the fragments range from 200 to 600bp. DDRT using total RNA produced 19 violet-and 17 white-specific cDNA fragments, ranging from 200 to 600bp. The fragments obtained by both AFLP and DDRT had been cloned into pGEM T-easy vector, amplified and subjected to the nucleotide sequence analyses. As a result of Blast sequence analysis, most of them sequenced up to date showed no similarity to any Known gene, while few has similarity to known animal or plant genes. An AFLP clone V6, for example, has a strong sequence similarity to the human transcription factor LZIP-alpha mRNA and a DDRT clone W19 to Solanum tuberosum 3-hydroxy-3-methylglutaryl coenzyme A reductase mRNA.

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Cloning and Characterization of Soybean IFS (Isoflavone Synthase) Genes from Korean Cultivar, Sinpaldalkong (신팔달콩 유래 IFS (isoflavone synthase)유전자 클로닝 및 기능 규명)

  • Park, Hayng-Mi;Shin, Sang-Hyun;Ko, Jong-Min;Yi, Gi-Hwan;Nam, Min-Hee;Chung, Young-Soo;Chung, Won-Bok;Lee, Jai-Heon;Park, Seong-Whan
    • Journal of Life Science
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    • v.14 no.1
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    • pp.38-44
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    • 2004
  • Two genes, SinIFS1 and SinIFS2 from Korean soybean cultivar, Sinpaldalkong known as one of isoflavonerich cultivars, were cloned with PCR and degenerate primers. The sequences of two genes were analyzed with previously reported IFS genes of leguminous plants and their expression pattern in various environmental conditions was surveyed. The genomic clone of SinIFS1 contained 1,828bp nucleotides and encoded a polypeptide of 521 amino acids, and 1912bp nucleotides and a polypeptide of 521 amino acids for SinIFS2. Both genes included several conserved motifs, oxygen binding and activation (A/G-G-X-E/D-T-T/S), ERR triad (E...R....R), and heme binding (F-X-X-G-X-R-X-C-X-G) domain, which are typical in any member of cytochrome P45O superfamily. Very high sequence homology (>98%) was observed in the comparison with other IFSs of legumes. In the northern blot analysis to check the expression and increase of SinIFS1 to various environmental renditions (low temperature, light, dark, UV, and fungal elicitor), the most significant induction, more than 6 times of transcript level compared to the dark treatment as a control, was observed from the fungal elicitor treatment. The next up-regulated expression was from UV treatment (4${\times}$), low temperature and light conditions.

Estimation of micro-biota in the Upo wetland using eukaryotic barcode molecular markers

  • Park, Hyun-Chul;Bae, Chang-Hwan;Jun, Ju-Min;Kwak, Myoung-Hai
    • Journal of Ecology and Environment
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    • v.34 no.3
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    • pp.323-331
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    • 2011
  • Biodiversity and the community composition of micro-eukaryotic organisms were investigated in the Upo wetland in Korea using molecular analysis. Molecular identification was performed using cytochrome oxidase I (COI) and small subunit ribosomal DNA (SSU rDNA). The genomic DNA was isolated directly from soil samples. The COI and SSU rDNA regions were amplified using universal primers and then sequenced after cloning. In a similarity search of the obtained sequences with BLAST in the Genbank database, the closely related sequences from NCBI were used to identify the amplified sequences. A total of six eukaryotic groups (Annelida, Arthropoda, Rotifera, Chlorophyta, Bacillariophyta, and Stramenopiles) with COI and six groups (Annelida, Arthropoda, Rotifera, Alveolata, Fungi, and Apicomplexa) with SSU rDNA genes were determined in the Upo wetland. Among 38 taxa in 20 genera, which are closely related to the amplified sequences, 10 genera (50%) were newly reported in Korea and five genera (25%) were shown to be distributed in the Upo wetland. This approach is applicable to the development of an efficient method for monitoring biodiversity without traditional taxonomic processes and is expected to produce more accurate results in depositing molecular barcode data in the near future.

Mycotoxins and Their Biotransformation in the Rumen: A Review

  • Upadhaya, Santi Devi;Park, M.A.;Ha, Jong-K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.9
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    • pp.1250-1260
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    • 2010
  • Mycotoxins are secondary metabolites produced by fungi. These toxins pose serious health concerns to animals as well as human beings. Biodegradation of these mycotoxins has been considered as one of the best strategies to decontaminate food and feedstuffs. Biodegradation employs the application of microbes or enzymes to contaminated food and feedstuffs. Ruminants are considered to be resistant to the adverse effects of mycotoxins presumably due to the biodegrading ability of rumen microbes compared to mono-gastric animals. Therefore, rumen microbial source or microbial enzyme could be a great asset in biological detoxification of mycotoxins. Isolation and characterization of pure culture of rumen microorganisms or isolation and cloning of genes encoding mycotoxin-degrading potential would prove to have overall beneficial impact in the food and feed industry.

Community Structures of Arbuscular Mycorrhizal Fungi in Soils and Plant Roots Inhabiting Abandoned Mines of Korea

  • Park, Hyeok;Lee, Eun-Hwa;Ka, Kang-Hyeon;Eom, Ahn-Heum
    • Mycobiology
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    • v.44 no.4
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    • pp.277-282
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    • 2016
  • In this study, we collected rhizosphere soils and root samples from a post-mining area and a natural forest area in Jecheon, Korea. We extracted spores of arbuscular mycorrhizal fungi (AMF) from rhizospheres, and then examined the sequences of 18S rDNA genes of the AMF from the collected roots of plants. We compared the AMF communities in the post-mining area and the natural forest area by sequence analysis of the AMF spores from soils and of the AMF clones from roots. Consequently, we confirmed that the structure of AMF communities varied between the post-mining area and the natural forest area and showed significant relationship with heavy metal contents in soils. These results suggest that heavy metal contamination by mining activity significantly affects the AMF community structure.

Taxonomic Studies of the Beta Hemolysis-causing Pathogen Bacillus cereus Isolated from Sea Water

  • Kim, Sam-Sun;Park, Yong-Ha;Lee, Jung-Sook;Yoon, Jung-Hoon;Shin, Yong-Kook;Rhee, In-Koo;Kim, Young-Jae
    • Journal of Microbiology and Biotechnology
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    • v.8 no.1
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    • pp.67-73
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    • 1998
  • A bacterial strain that excretes hemolysins and proteases into the growth medium was isolated from sea water and designated as KYJ 961. A nearly complete nucleotide sequence of a 16S ribosomal RNA gene from the isolate was determined following the isolation and cloning of amplified genes. On the basis of the 16S ribosomal DNA sequence data, and morphological, chemotaxonomic, and physiological characteristics, strain KYJ 961 was classified as a strain of Bacillus cereus.

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High Level of Bacterial Diversity and Novel Taxa in Continental Shelf Sediment

  • Hong, Jin-Kyung;Cho, Jae-Chang
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.771-779
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    • 2012
  • The bacterial diversity of the continental shelf sediment in the Yellow Sea was investigated by the cloning and sequencing of PCR-amplified 16S rRNA genes. The majority of the cloned sequences were distinct phylotypes that were novel at the species level. The richness estimator indicated that the sediment sample might harbor up to 32 phylum-level taxa. A large number of low-abundance, phylum-level taxa accounted for most of the observed phylogenetic diversity at our study site, suggesting that these low-abundance taxa might play crucial roles in the shelf sediment ecosystem.

Identification of Tumor Suppressor Genes on Chromosome 21 (제21번 염색체의 종양억제유전자 발굴)

  • Lee, Eung-Bae;Choi, Jin-Eun;Jang, Jin-Sung;Park, Jae-Yong
    • Journal of Chest Surgery
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    • v.42 no.2
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    • pp.141-147
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    • 2009
  • Background: We performed this study to identify the tumor suppressor genes located in the long arm of chromosome 21 in non-small cell lung cancer. Material and Method: The genes of USP25 in 21q11.2, NCAM2, ADAMTS1 in 21q21.2, and Claudin-8 (CLDN8), Claudin-17 (CLDN17) and TIAM1 in 21q22.1 were investigated for their gene expressions, genetic alterations and promoter methylation. Result: The expressions of CLDN8 and CLDN17 were significantly decreased in 7 (L132, H157, H358, H522, H1299, H1703 and HCC2108) of 13 cell lines, and the expression of ADAMTS1 was also significantly reduced in 6 cell lines (A549, SW900, H1299, H1373, H1703 and H1793). There were no genetic alterations by PCR-SSCP and cDNA cloning in the cell lines with a decreased gene. In the cell lines with a decreased gene expression, the mRNA expression was increased significantly with treatment of 5-Aza-CdR. Conclusion: These results suggest that the ADMTS1, CLDN8 and CLDN17 may act as tumor suppressor genes.

Cloning and Sequence Analysis of the Aminoglycoside Resistance Gene from a Nebramycin Complex Producer, Streptoalloteichus hindustanus

  • Hyun, Chang-Gu;Kim, Jong-Woo;Han, Jae-Jin;Choi, Young-Nae;Suh, Joo-Won
    • Journal of Microbiology and Biotechnology
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    • v.8 no.2
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    • pp.146-151
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    • 1998
  • The aminoglycoside multiple-resistance determinant from Streptoalloteichus hindustanus was cloned into Streptomyces lividans and named nbrB. The 1.2-kb ApaI- BclI fragment encompassing nbrB was located within a 2.6-kb ApaI fragment by successive subcloning experiments. The complete DNA nucleotide sequence of 1.2-kb containing nbrB was determined. The sequence contains an open reading frame that putatively encodes a polypeptide of 281 amino acids with a predicted molecular weight of 30,992. The deduced amino acid sequence of nbrB shows identities of 85.1% to kgmB of S. tenebrarius, 59.6% to sgm of Micromonospora zionensis, and 57.7% to grm of M. rosea. The similarity of nbrB to kgmB suggests that nbrB encodes a 16S rRNA methylase similar to that encoded by kgmB and that both genes might be derived from a common ancestral gene.

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