• Title/Summary/Keyword: Gene Technology

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Cloning and Expression of Phytochelatin Synthase 1 Gene from Rhizophora stylosa Exposed to Cadmium and Copper (카드뮴과 구리에 노출된 Rhizophora stylosa 의 phytochelatin synthase 1 유전자 클로닝 및 발현)

  • Lee, Gunsup;Hwang, Jinik;Park, Mirye;Chung, Youngjae;Lee, Taek-Kyun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.6
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    • pp.3114-3119
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    • 2013
  • The mangrove ecosystems have the capacity to act as a sink of heavy metals entering aquatic ecosystems. Despite their potential exposure to metal contaminated sediments, mangroves appear to be highly tolerant to heavy metals. In this study, we cloned metal tolerance gene from mangrove plant. Using CTAB method, RNA were isolated from leaves and root tissue of Rhizophora stylosa habitated at Weno island in Micronesia Chuuk lagoon using CTAB method and phytochelatin synthase 1 (PCS1) gene was cloned using gene specific primers. Expression of PCS1 gene was increased 1.91 fold and 2.72 fold in mangrove propagules exposed to 100 ppb Cd and 10 ppb Cu, respectively. These results indicate that expression of PCS1 gene are promising tools for health assessment of mangrove ecosystem.

Isolation of a Leucoanthocyanidin Dioxygenase (LDOX) Gene from a Spray-type Chrysanthemum (Dendranthema × grandiflorum) and Its Colored Mutants (스프레이형 국화와 화색변이체로부터 Leucoanthocyanidin dioxygenase (LDOX) 유전자의 분리)

  • Chung, Sung-Jin;Lee, Geung-Joo;Lee, Hye-Jung;Kim, Jin-Baek;Kim, Dong-Sub;Kang, Si-Yong
    • Horticultural Science & Technology
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    • v.28 no.5
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    • pp.818-827
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    • 2010
  • A full-length cDNA and genomic DNA of a $leucoanthocyanidin$ $dioxygenase$ ($DgLDOX$) gene was isolated from the petals of chrysanthemum 'Argus', and comparative features of the gene among three flower color mutants derived from a gamma-ray mutagenesis were characterized. The cDNA coding region of the gene was 1068 bp and was translated into 356 amino acids accordingly. The genomic DNA size was 1346 bp for 'Argus', while three mutants revealed ranges of 1363 to 1374 bp. A single intron between two coding exons for the $DgLDOX$ gene was found, of which size was 112 bp for 'Argus', but 128 or 137 bp for three flower color mutants, indicating that a genomic insertion in the intron occurred during the gamma-ray mutagenesis. DNA blot analysis revealed the $DgLDOX$ gene presenting as a single copy in the chrysanthemum genome. The $DgLDOX$ gene was expressed in both 'Argus' of light-pink color and two purple color mutants (AM1 and AM3) but had very weak expression in only white color mutant (AM2). The results demonstrated that variations in the flower color of the mutants might be associated with changes in the amino acid moieties in the coding exons or fragment insertions in the intron of the $DgLDOX$ gene, which potentially resulted in less expression of the gene in the white colored mutant.

Cloning and Iron Transportation of Nucleotide Binding Domain of Cryptosporidium andersoni ATP-Binding Cassette (CaABC) Gene

  • Wang, Ju-Hua;Xue, Xiu-Heng;Zhou, Jie;Fan, Cai-Yun;Xie, Qian-Qian;Wang, Pan
    • Parasites, Hosts and Diseases
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    • v.53 no.3
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    • pp.335-339
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    • 2015
  • Cryptosporidium andersoni ATP-binding cassette (CaABC) is an important membrane protein involved in substrate transport across the membrane. In this research, the nucleotide binding domain (NBD) of CaABC gene was amplified by PCR, and the eukaryotic expression vector of pEGFP-C1-CaNBD was reconstructed. Then, the recombinant plasmid of pEGFP-C1-CaNBD was transformed into the mouse intestinal epithelial cells (IECs) to study the iron transportation function of CaABC. The results indicated that NBD region of CaABC gene can significantly elevate the transport efficiency of $Ca^{2+}$, $Mg^{2+}$, $K^+$, and $HCO_3{^-}$ in IECs (P<0.05). The significance of this study is to find the ATPase inhibitors for NBD region of CaABC gene and to inhibit ATP binding and nutrient transport of CaABC transporter. Thus, C. andersoni will be killed by inhibition of nutrient uptake. This will open up a new way for treatment of cryptosporidiosis.

Cloning and Characterization of the Tetracycline Resistant Gene, tetB, from Vibrio parahaemolyticus

  • Kang, Min-Seung;Park, Kun-Ba-Wui;Hwang, Hye-Jin;Bae, Hyang-Nam;Lim, Keun-Sik;Eom, Sung-Hwan;Kim, Young-Mog
    • Fisheries and Aquatic Sciences
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    • v.12 no.1
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    • pp.24-28
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    • 2009
  • A tetracycline resistant Vibrio parahaemolyticus, capable of growing on TCBS medium containing tetracycline, was isolated from cultivated fishes. A gene responsible for the tetracycline resistance was cloned from chromosomal DNA of the V. parahaemolyticus strain using Escherichia coli KAM3, which lacks major multi-drug efflux pumps (${\Delta}acrB$) as host cells. The nucleotide sequence and homology analysis revealed an open reading frame (ORF) for tetracycline resistance protein (TetB). In order to characterize the antibiotic resistance of TetB originated from the V. parahaemolyticus strain, the gene was sub cloned into plasmid pSTV28. The resulting plasmid was designated as pSTVTetB and transformated into E. coli KAM3. E. coli KAM3 cells harboring the recombinant plasmid pSTVTetB are able to grow on plates containing tetracycline and oxytetracycline but not doxycycline, indicating that the tetB gene confers the tetracycline- and oxytetracycline-resistance to the host cell.

Complex Regulatory Network of MicroRNAs, Transcription Factors, Gene Alterations in Adrenocortical Cancer

  • Zhang, Bo;Xu, Zhi-Wen;Wang, Kun-Hao;Lu, Tian-Cheng;Du, Ye
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.4
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    • pp.2265-2268
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    • 2013
  • Several lines of evidence indicate that cancer is a multistep process. To survey the mechanisms involving gene alteration and miRNAs in adrenocortical cancer, we focused on transcriptional factors as a point of penetration to build a regulatory network. We derived three level networks: differentially expressed; related; and global. A topology network ws then set up for development of adrenocortical cancer. In this network, we found that some pathways with differentially expressed elements (genetic and miRNA) showed some self-adaption relations, such as EGFR. The differentially expressed elements partially uncovered mechanistic changes for adrenocortical cancer which should guide medical researchers to further achieve pertinent research.

Studies on the $HIS_5$ Gene of Yeast -$HIS_5-lacZ$ fusion and regulation in Saccharomyces cerevisiae- (효모 $HIS_5$ 유전자에 관한 연구 -Saccharomyces cerevisiae의 $HIS_5-lacZ$ 융합과 조절-)

  • Chung, Dong-Hyo;Oshima, Yasuji;Nishiwaki, Kyoni
    • Applied Biological Chemistry
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    • v.28 no.1
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    • pp.36-47
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    • 1985
  • $HIS_5$ gene of Saccaromyces cerevisiae was cloned into pBR 322 and also into pSH 610 shuttle vector. $HIS_5$ gene was expressed as promoter of lactose operon(lacZ). And $HIS_5-lacZ$ fusion was intergrated into chromosome III of yeast. $HIS_5$ gene in yeast growth was controlled general amino acid control mechanism.

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Identification and Characterization of the wbpO Gene Essential for Lipopolysaccharide Synthesis in Vibrio vulnificus

  • Park Na-Young;Lee Jeong-Hyun;Lee Byung-Cheol;Kim Tae-Sung;Choi Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.16 no.5
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    • pp.808-816
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    • 2006
  • A wbpO gene encoding a putative UDP-N-acetylo-galactosamine dehydrogenase was identified and cloned from Vibrio vulnificus. LPS production was altered by disruption of the wbpO gene through allelic exchanges. The function of the wbpO gene in virulence assessed in vitro and in mice revealed that WbpO is important in both the pathogenesis of V. vulnificus and the biosynthesis of LPS.

A Partial Nucleotide Sequence of Chitin Synthase (CHS) Gene from Rice Blast Fungus, Pyricularia oryzae and Its Cloning

  • Hwang, Cher-Won;Park, In-Cheol;Yeh, Wan-Hae;Takagi, Masamchi;Ryu, Jin-Chang
    • Journal of Microbiology and Biotechnology
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    • v.7 no.2
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    • pp.157-159
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    • 1997
  • A 340-bp chitin synthase gene(CHS) fragment was cloned from the genomic DNA of Pyricularia oryzae using a PCR process with two primer DNAs corresponding to highly conserved sequences within fungal CHS genes. The entire DNA nucleotide sequences of the cloned DNA fragment were determined and analyzed. The amino acid sequences deduced from the nucleotide sequence of the amplified DNA fragment showed 86% homology to that of the Aspergillus fumigatus CHSE gene (9). Using this PCR-amplified DNA, about 2.3 kb of including the PCR fragment of CHSE gene was cloned from genomic library.

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Cloning of Cellobiohydrolase Gene (cbhI) in Radiation Induced Mutant of Pleurotus florida

  • Lee, Young-Keun;Sathesh-Prabu, Chandran;Kim, Min Kyung
    • Journal of Radiation Industry
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    • v.8 no.2
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    • pp.65-69
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    • 2014
  • The cellobiohydrolase gene (cbhI), a key component of a cellulolytic system, of a mutant PfCM4 (Pleurotus florida), developed through gamma ray radiation mutagenesis, was isolated and cloned. The deduced amino acid sequence was closely related to the glycoside hydrolase family 7 (GH7). The molecular weight of the deduced amino acid sequence of cbhI gene was found to be 22.4 kDa. Though the percent identity was found to be much less (35.61%) between the wild type and mutant, the cellulolytic activity of PfCM4 was 17.24% higher than that of the wild type. This shows that the catalytic domain of the cbhI gene was conserved in the mutant PfCM4.

Characterization of Structural Variations in the Context of 3D Chromatin Structure

  • Kim, Kyukwang;Eom, Junghyun;Jung, Inkyung
    • Molecules and Cells
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    • v.42 no.7
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    • pp.512-522
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    • 2019
  • Chromosomes located in the nucleus form discrete units of genetic material composed of DNA and protein complexes. The genetic information is encoded in linear DNA sequences, but its interpretation requires an understanding of three-dimensional (3D) structure of the chromosome, in which distant DNA sequences can be juxtaposed by highly condensed chromatin packing in the space of nucleus to precisely control gene expression. Recent technological innovations in exploring higher-order chromatin structure have uncovered organizational principles of the 3D genome and its various biological implications. Very recently, it has been reported that large-scale genomic variations may disrupt higher-order chromatin organization and as a consequence, greatly contribute to disease-specific gene regulation for a range of human diseases. Here, we review recent developments in studying the effect of structural variation in gene regulation, and the detection and the interpretation of structural variations in the context of 3D chromatin structure.