• Title/Summary/Keyword: Arabidopsis thaliana

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Isolation and Characterization of Bacillus cereus A-139 Producing Auxin from East Coast Sand Dunes (동해안 사구로부터 Auxin을 생산하는 Bacillus cereus A-139의 분리 및 그 특성)

  • So, Jai-Hyun;Kim, Duk-Jin;Shin, Jae-Ho;Yu, Choon-Bal;Rhee, In-Koo
    • Korean Journal of Environmental Agriculture
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    • v.28 no.4
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    • pp.447-452
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    • 2009
  • A bacterium, which was named to be Bacillus cereus A-139, secreting auxin was isolated from the east coast sand dunes in Korea. The secretion of auxin was confirmed by HPLC. When cultured in LB broth, Bacillus cereus A-139 produced $16.12\;{\mu}$g/mL auxin after 8 days in LB broth. Bacillus cereus A-139 produced $49\;{\mu}$g/mL auxin and $162.6\;{\mu}$g/mL by the addition of 2% tryptone and 0.1% tryptophan, respectively. The root growth of Arabidopsis thaliana was retarded by Bacillus cereus A-139 culture broth up to 57% but the formation of lateral roots was increased up to almost twice after 4 days incubation. Also the formation of lateral roots of mung bean was increased up to 57% after 10 days incubation.

Structure and Expression of a Perilla (Perilla frutescens Britt) Gene, PfFAD3, Encoding the Microsomal ${\omega}-3$ Fatty Acid Desaturase

  • Lee, Hyang-Hwa;Pyee, Jae-Ho
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.5
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    • pp.424-427
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    • 2004
  • Microsomal ${\omega}-3$ fatty acid desaturase (FAD3) is an essential enzyme in the production of the n-3 polyunsaturated fatty acid ${\alpha}-linolenic$ acid during the seed developing stage. To understand the regulatory mechanism of the gene encoding the ${\omega}-3$ fatty acid desaturase, a genomic fragment corresponding to the previously isolated perilla seed PfFAD3 cDNA was amplified from perilla (Perilla frutescens Britt) by GenomeWalker PCR. Sequence analysis of the fragment provided with identification of a 1485-bp 5'-upstream region and a 241-bp intron in the open reading frame. To determine the tissue-specificity of the PfFAD3 gene expression, the 5'-upstream region was fused to the ${\beta}-glucuronidase$ (GUS) gene and incorporated into Arabidopsis thaliana. Histochemical assay of the transgenic plants showed that GUS expression was restricted to seed and pollen, showing that PfFAD3 gene was exclusively expressed in those tissues.

Identification and Isolation of Differentially Expressed Gene in Response to Cold Stress in a Green Alga, Spirogyra varians (Zygnematales)

  • Han, Jong-Won;Yoon, Min-Chul;Lee, Key-Pyoung;Kim, Gwang-Hoon
    • ALGAE
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    • v.22 no.2
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    • pp.131-139
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    • 2007
  • The expression of genes responding to cold stress in a freshwater alga, Spirogyra varians, was studied by using differential expression gene (DEG) method. A gene strongly up-regulated in 4°C was isolated and designated as SVCR2 (Spirogyra varians cold regulated) gene. The cDNA encoding SVCR2 was cloned using λZAP cDNA library of Spirogyra varians. The deduced amino acid had a sequence similarity with trans-membrane protein in Arabidopsis thaliana (Q9M2D2, 52.7%). Northern blot analysis demonstrated that transcript level of SVCR2 increased about 10 fold under low temperature (4°C), compared with that cultured at warm (20°C) conditions. The expression of SVCR2 was also affected by light conditions. When the plants were exposed to high light (HL) (1200 μmol photon m–2 s–1), the expression of SVCR2 began within 2 hrs. This gene expression lasted for 4 hrs and decreased afterwards. Under the blue light (470 nm) condition, the expression of this gene was induced in same way as HL treatment, even under less than 100 μmol photon m–2 s–1. But red light (650 nm) and UV-A irradiation did not affect the expression of SVCR2.

Isolation and characterization of BrMDR1 a novel MDR-type ATP-binding cassette (ABC) transporter in Brassica rapa L.

  • Lee, Sun-Yong;Jung, Yu-Jin;Kang, Kwon-Kyoo
    • Korean Journal of Plant Resources
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    • v.22 no.3
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    • pp.273-280
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    • 2009
  • A cDNA clone encoding a MDR-like ABC transporter protein was isolated from Brassica rapa seedlings, through rapid amplification of cDNA ends (RACE). This gene (named as Brmdr 1; GenBank accession no.: DQ296184 ) had a total length of 4222 bp with an open reading frame of 3900 bp, and encoded a predicted polypeptide of 1300 amino acids with a molecular weight of 143.1 kDa. The BrMDR1 protein shared 71.0, 62.5, 60.0 and 58.2% identity with other MDR proteins isolated from Arabidopsis thaliana (AAN28720), Coptis japonica (CjMDR), Gossypium hirsutum (GhMDR) and Triticum aestivum (TaMDR) at amino acid level, respectively. Southern blot analysis showed that Brmdr1 was a low-copy gene. Expression pattern analysis revealed that Brmdr1 constitutively expressed in the root, stem petals and stamens, but with lower expression in leaves and open flowers. The domains analysis showed that BrMDR1 protein possessed two transmembrane domains (TMDs) and two nucleotide binding domains (NBDs) arranging in "TMD1-NBD1-TMD2-NBD2" direction, which is consistent with other MDR transporters. Within NBDs three characteristic motifs common to all ABC transporters, "Walker A", "Walker B" and C motif, were found. These results indicate that BrMDR1 is a MDR-like ABC transporter protein that may be involved in the transport and accumulation of secondary metabolites.

Current status and prospects of the meiosis-specific function of recombinase in plants (식물의 감수분열에서 상동 재조합 효소 특이 기능의 연구현황 및 전망)

  • Jung, Yu Jin;Nam, Ki Hong;Kim, Tae Sung;Lee, In Hae;Cho, Yong-Gu;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.45 no.1
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    • pp.1-8
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    • 2018
  • Meiosis is a specialized cell division, essential in most reproducing organisms to halve the number of chromosomes, thereby enabling the restoration of ploidy levels during fertilization. A key step in meiosis is homologous recombination, which promotes homologous pairing and generates crossovers (COs) to connect homologous chromosomes until their separation at anaphase I. These CO sites, seen cytologically as chiasmata, represent a reciprocal exchange of genetic information between two homologous non-sister chromatids. RAD51, the eukaryotic homolog of the bacterial RecA recombinase, plays a central role in homologous recombination (HR) in yeast and animals. Loss of RAD51 function causes lethality in the flowering plant, Arabidopsis thaliana, suggesting that RAD51 has a meiotic stage-specific function that is different from homologous pairing activity.

A transcription factor "OsNAC075" is essential for salt resistance in rice (Oryza sativa L.)

  • Jung, Yu-Jin;Lee, Myung-Chul;Kang, Kwon-Kyoo
    • Journal of Plant Biotechnology
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    • v.38 no.1
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    • pp.94-104
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    • 2011
  • Salt stress is a major environmental factor influencing plant growth and development. To identify salt tolerance determinants, we systematically screened salt sensitive rice mutants by use of the Activator/Dissociation (Ac/Ds) transposon tagging system. In this study, we focused on the salt sensitive mutant line, designated SSM-1. A gene encoding a NAC transcription factor homologue was disrupted by the insertion of a Ds transposon into SSM-1 line. The OsNAC075 gene (EU541472) has 7 exons and encodes a protein (486-aa) containing the NAC domain in its N-terminal region. Sequence comparison showed that the OsNAC075 protein had a strikingly conserved region at the N-terminus, which is considered as the characteristic of the NAC protein family. OsNAC075 protein was orthologous to Arabidopsis thaliana ANAC075. Phylogenetic analysis confirmed OsNAC075 belonged to the OsNAC3 subfamily, which plays an important role in response to stress stimuli. RT-PCR analysis showed that the expression of OsNAC075 gene was rapidly and strongly induced by stresses such as NaCl, ABA and low temperature ($4^{\circ}C$). Our data suggest that OsNAC075 holds promising utility in improving salt tolerance in rice.

Metabolic Engineering of Escherichia coli for the Biological Synthesis of 7-O-Xylosyl Naringenin

  • Simkhada, Dinesh;Kim, EuiMin;Lee, Hei Chan;Sohng, Jae Kyung
    • Molecules and Cells
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    • v.28 no.4
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    • pp.397-401
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    • 2009
  • Flavonoids are a group of polyphenolic compounds that have been recognized as important due to their physiological and pharmacological roles and their health benefits. Glycosylation of flavonoids has a wide range of effects on flavonoid solubility, stability, and bioavailability. We previously generated the E. coli BL21 (DE3) ${\Delta}pgi$ host by deleting the glucose-phosphate isomerase (Pgi) gene in E. coli BL21 (DE3). This host was further engineered for whole-cell biotransformation by integration of galU from E. coli K12, and expression of calS8 (UDP-glucose dehydrogenase) and calS9 (UDP-glucuronic acid decarboxylase) from Micromonospora echinospora spp. calichensis and arGt-4 (7-O-glycosyltransferase) from Arabidopsis thaliana to form E. coli (US89Gt-4), which is expected to produce glycosylated flavonoids. To test the designed system, the engineered host was fed with naringenin as a substrate, and naringenin 7-O-xyloside, a glycosylated naringenin product, was detected. Product was verified by HPLC-LC/MS and ESI-MS/MS analyses. The reconstructed host can be applied for the production of various classes of glycosylated flavonoids.

Caffeoyl Shikimate Esterase has a Role in Endocarp Lignification in Peach (Prunus persica L.) Fruit

  • Liu, Jinyi;Hu, Xiao;Yu, Jia;Yang, Aizhen;Liu, Yueping
    • Horticultural Science & Technology
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    • v.35 no.1
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    • pp.59-68
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    • 2017
  • Caffeoyl shikimate esterase (CSE) is a key enzyme in lignin synthesis in Arabidopsis thaliana. To determine the role of CSE in lignification of the endocarp in peach (Prunus persica L.) fruit, we cloned and characterized the P. persica CSE homolog, which we designated PpCSE. The 954 - bp PpCSE gene encoded a 317 - amino acid polypeptide. PpCSE expression patterns in the mesocarp and endocarp changed during peach fruit development. There was no significant difference between the expression levels of PpCSE in the mesocarp and endocarp at 39 and 44 days after full bloom (DAFB), but the expression level of PpCSE in the endocarp at 50 and 55 DAFB was 80.73 and 72.75 times higher, respectively, than that in the mesocarp. During peach fruit development, PpCSE expression in the endocarp increased rapidly; the relative PpCSE expression level at 50 DAFB was 122.70 times higher than that at 39 DAFB. At the protein level, CSE was detected in the peach fruit endocarp at 50 and 55 DAFB. Our study suggests that PpCSE expression during peach fruit development is closely related to the degree of endocarp lignification.

A Gene Encoding $\beta$-amylase from Saprolegnia parasitica and Its Expression in Saccharomyces cerevisiae

  • Kim, Hee-Ok;Park, Jeong-Nam;Shin, Dong-Jun;Lee, HwangHee Blaise;Chun, Soon-Bai;Bai, Suk
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.529-533
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    • 2001
  • The ${\beta}$-Amylase cDNA fragment from the oomcete Saprolegnia parasitica was cloned by reverse transcription-polymerase chain reaction (RT-PCR) using degenerate oligonucleotide primers derived from conserved ${\beta}$-amylase sequences. The 5'and 3'regions of the $\beta$-amylase gene were amplified using the rapid amplification of cDNA ends (rACE) system. It consisted of an open reading frame of 1,350 bp for a protein of 450 amino acids. Comparison between the genomic and cDNA sequences revealed that the intron was not present in the coding region. The deduced amino acid sequence of the ${\beta}$-amylase gene had a 97% similarity to the ${\beta}$-amylase of Saprolegnia ferax, followed by 41% similarity to those of Arabidopsis thaliana, Hordeum vulgare, and Zea mays. The ${\beta}$-amylase gene was also expressed in Saccharomyces cerevisiae by placing it under the control of the alcohol dehydrogenase gene (ADC1) promoter.

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K-mer Based RNA-seq Read Distribution Method For Accelerating De Novo Transcriptome Assembly

  • Kwon, Hwijun;Jung, Inuk
    • Journal of the Korea Society of Computer and Information
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    • v.25 no.8
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    • pp.1-8
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    • 2020
  • In this paper, we propose a gene family based RNA-seq read distribution method in means to accelerate the overal transcriptome assembly computation time. To measure the performance of our transcriptome sequence data distribution method, we evaluated the performance by testing four types of data sets of the Arabidopsis thaliana genome (Whole Unclassified Reads, Family-Classified Reads, Model-Classified Reads, and Randomly Classified Reads). As a result of de novo transcript assembly in distributed nodes using model classification data, the generated gene contigs matched 95% compared to the contig generated by WUR, and the execution time was reduced by 4.2 times compared to a single node environment using the same resources.