• Title/Summary/Keyword: Gene characterization

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Characterization of a Squalene Synthase from the Thraustochytrid Microalga Aurantiochytrium sp. KRS101

  • Hong, Won-Kyung;Heo, Sun-Yeon;Park, Hye-Mi;Kim, Chul Ho;Sohn, Jung-Hoon;Kondo, Akihiko;Seo, Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.759-765
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    • 2013

  • The gene encoding squalene synthase (SQS) of the lipid-producing heterotrophic microalga Aurantiochytrium sp. KRS101 was cloned and characterized. The krsSQS gene is 1,551 bp in length and has two exons and one intron. The open reading frame of the gene is 1,164 bp in length, yielding a polypeptide of 387 predicted amino acid residues with a molecular mass of 42.7 kDa. The deduced krsSQS sequence shares at least four conserved regions known to be required for SQS enzymatic activity in other species. The protein, tagged with $His_6$, was expressed into soluble form in Escherichia coli. The purified protein catalyzed the conversion of farnesyl diphosphate to squalene in the presence of NADPH and $Mg^{2+}$. This is the first report on the characterization of an SQS from a Thraustochytrid microalga.

  • https://doi.org/10.4014/jmb.1212.12023 인용 PDF KSCI

Molecular Characterization of Silicon (Si) Transporter Genes, Insights into Si-acquisition Status, Plant Growth, Development, and Yield in Alfalfa

  • Md Atikur Rahman;Sang-Hoon Lee;Yowook Song;Hyung Soo Park;Jae Hoon Woo;Bo Ram Choi;Ki-Won Lee
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.43 no.3
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    • pp.168-176
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    • 2023

  • Silicon (Si) has the potential to improve plant growth and stress tolerance. The study aimed to explore Si-involving plant responses and molecular characterization of different Si-responsive genes in alfalfa. In this study, the exogenous supplementation of Si enhanced plant growth, and biomass yield. Si-acquisition in alfalfa root and shoot was higher in Si-supplemented compared to silicon deficient (-Si) plants, implying Si-acquisition has beneficial on alfalfa plants. As a consequence, the quantum efficiency of photosystem II (Fv/Fm) was significantly increased in silicon-sufficient (+Si) plants. The quantitative gene expression analysis exhibited a significant upregulation of the Lsi1, Lsi2, Lsi3, NIP5;1, and NIP6;1 genes in alfalfa roots, while BOR1, BOR4, NIP2, and NIP3 showed no significant variation in their expression. The MEME results further noticed the association of four motifs related to the major intrinsic protein (MIP). The interaction analysis revealed that NIP5;1 and Lsi1 showed a shared gene network with NIP2, BOR1, and BOR4, and Lsi2, Lsi3 and NIP3-1, respectively. These results suggest that members of the major intrinsic proteins (MIPs) family especially Lsi1, Lsi2, Lsi3, NIP5;1, and NIP6;1 genes helped to pass water and other neutral solutes through the cell membrane and those played significant roles in Si uptake and transport in plants. Together, these insights might be useful for alfalfa breeding and genome editing approaches for alfalfa improvement.

  • https://doi.org/10.5333/KGFS.2023.43.3.168 인용 PDF

Isolation and Characterization of hrp2+ Gene Related to SNF2 Family In Yeast (Schizosaccharomyces pombe에서 SNF2에 속하는 hrp2+ 유전자의 특성 연구)

  • Choi In Soon
    • Journal of Life Science
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    • v.15 no.2 s.69
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    • pp.192-196
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    • 2005

  • The SNF2/SW12 family comprises proteins from a variety of species with in vivo functions, such as transcriptional regulation, maintenance of chromosome stability during mitosis, and various types of DNA repair. This study was shown the characterization of hrp2+ gene which was isolated by PCR amplification using the conserved domain of SNF2 motifs. Sequence analysis of hrp2+ gene showed striking evolutionary conservation among the SNF2 family of proteins. The transcript of hrp2+ gene was found to be a 4.7 kb as identified by Northern hybridization. To investigate the inducibility of hrp2+ gene, transcript levels were examined after treating the cells to various DNA damaging agents. The transcripts of hrp2+ were induced by UV-irradiation. But the transcripts were not induced by treatment of $ 0.25\%$ Methylmethane sulfonate (MMS). These results implied that the effects of damaging agents are complex and different regulatory pathways exist for the induction of this gene. Hrp2 protein was purified near homogeneity by combination of affinity chromatography. We tested the purified Hrp2 protein for the helicase activity in an oligonucleotide release assay. However we were unable to detect any helicase activity associated with the Hrp2 protein, indicating that the helicase motifs in Hrp2 are merely indicators of a broader DNA-dependent ATPase activity.

  • https://doi.org/10.5352/JLS.2005.15.2.192 인용 PDF KSCI