• Title/Summary/Keyword: Mutant

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Inhibitory Effect of the Branch Extracts from Taxillus yadoriki Parasitic to Neolitsea sericea against the Cell Proliferation in Human Lung Cancer Cells, A549 (참식나무(Neolitsea sericea) 기주 참나무겨우살이(Taxillus yadoriki) 가지 추출물의 폐암세포 A549에 대한 세포생육 억제활성)

  • Park, Su Bin;Kim, Ha Na;Kim, Jeong Dong;Park, Gwang Hun;Eo, Hyun Ji;An, Mi-Yun;Jeong, Jin Boo
    • Korean Journal of Plant Resources
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    • v.32 no.2
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    • pp.109-115
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    • 2019
  • In this study, we evaluated the anti-cancer activity and potential molecular mechanism of 70% ethanol extracts of branches from Taxillus yadoriki parasitic to Neolitsea sericea (TN-NS-B) against human lung cancer cells, A549. TY-NS-B dose-dependently suppressed the growth of A549 cells. TY-NS-B decreased ${\beta}$-catenin protein level, but not mRNA level in A549 cells. The downregulation of ${\beta}$-catenin protein level by TY-NS-B was attenuated in the presence of MG132. Although TY-NS-B phosphorylated ${\beta}$-catenin protein, the inhibition of $GSK3{\beta}$ by LiCl did not blocked the reduction of ${\beta}$-catenin by TY-NS-B. In addition, TY-NS-B decreased ${\beta}$-catenin protein in A549 cells transfected with Flag-tagged wild type ${\beta}$-catenin or Flag-tagged S33/S37/T41 mutant ${\beta}$-catenin construct. Our results suggested that TN-NS-B may downregulate ${\beta}$-catenin protein level independent on $GSK3{\beta}$-induced ${\beta}$-catenin phosphorylation. Based on these findings, TY-NS-B may be a potential candidate for the development of chemopreventive or therapeutic agents for human lung cancer.

Ex Vivo Raman Spectroscopy Measurement of a Mouse Model of Alzheimer's Disease (라만 기반 치매 모델의 뇌조직 분광 특성 측정)

  • Ko, Kwanhwi;Seo, Younghee;Im, Seongmin;Lee, Hongki;Park, Ji Young;Chang, Won Seok;Kim, Donghyun
    • Korean Journal of Optics and Photonics
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    • v.33 no.6
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    • pp.331-337
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    • 2022
  • Raman spectroscopy is an optical technique that can identify molecules in a label-free manner, and is therefore heavily investigated in various areas ranging from biomedical engineering to materials science. Probe-based Raman spectroscopy can perform minimally invasive chemical analysis, and thus has potential as a real-time diagnostic tool during surgery. In this study, Raman experimentation was calibrated by examining the Raman shifts with respect to the concentrations of chemical substances. Raman signal characteristics, targeted for normal mice and cerebral tissues of the 5xFAD dementia mutant model with accumulated amyloid beta plaques, were measured and analyzed to explore the possibility of diagnosis of Alzheimer's disease. The application to the diagnosis of dementia was cross-validated by measuring Raman signals of amyloid beta. The results suggest the potential of Raman spectroscopy as a diagnostic tool that may be useful in various areas of application.

Translocalization of enhanced PKM2 protein into the nucleus induced by cancer upregulated gene 2 confers cancer stem cell-like phenotypes

  • Yawut, Natpaphan;Kaowinn, Sirichat;Cho, Il-Rae;Budluang, Phatcharaporn;Kim, Seonghye;Kim, Suhkmann;Youn, So Eun;Koh, Sang Seok;Chung, Young-Hwa
    • BMB Reports
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    • v.55 no.2
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    • pp.98-103
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    • 2022
  • Increased mRNA levels of cancer upregulated gene (CUG)2 have been detected in many different tumor tissues using Affymetrix microarray. Oncogenic capability of the CUG2 gene has been further reported. However, the mechanism by which CUG2 overexpression promotes cancer stem cell (CSC)-like phenotypes remains unknown. With recent studies showing that pyruvate kinase muscle 2 (PKM2) is overexpressed in clinical tissues from gastric, lung, and cervical cancer patients, we hypothesized that PKM2 might play an important role in CSC-like phenotypes caused by CUG2 overexpression. The present study revealed that PKM2 protein levels and translocation of PKM2 into the nucleus were enhanced in CUG2-overexpressing lung carcinoma A549 and immortalized bronchial BEAS-2B cells than in control cells. Expression levels of c-Myc, CyclinD1, and PKM2 were increased in CUG2-overexpressing cells than in control cells. Furthermore, EGFR and ERK inhibitors as well as suppression of Yap1 and NEK2 expression reduced PKM2 protein levels. Interestingly, knockdown of β-catenin expression failed to reduce PKM2 protein levels. Furthermore, reduction of PKM2 expression with its siRNA hindered CSC-like phenotypes such as faster wound healing, aggressive transwell migration, and increased size/number of sphere formation. The introduction of mutant S37A PKM2-green fluorescence protein (GFP) into cells without ability to move to the nucleus did not confer CSC-like phenotypes, whereas forced expression of wild-type PKM2 promoted such phenotypes. Overall, CUG2-induced increase in the expression of nuclear PKM2 contributes to CSC-like phenotypes by upregulating c-Myc and CyclinD1 as a co-activator.

Production of Carotenoids by Bacteria; Carotenoid Productivity and Availability (박테리아에 의한 카로티노이드 생산; 카로티노이드 생산성 및 활용 가능성)

  • Choi, Seong Seok;Kim, Gun-Do
    • Journal of Life Science
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    • v.32 no.5
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    • pp.411-419
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    • 2022
  • Carotenoids are red, orange, and yellow fat-soluble pigments that exist in nature, and are known as physiologically active substances with various functions, such as provitamin A, antioxidant, anti-inflammatory, and anticancer. Because of their physiological activity and color availability, carotenoids are widely used in the food, cosmetics, and aquaculture industries. Currently, most carotenoids used industrially use chemical synthesis because of their low production cost, but natural carotenoids are in the spotlight because of their safety and physiologically active effects. However, the production of carotenoids in plants and animals is limited for economic reasons. Carotenoids produced by bacteria have a good advantage in replacing carotenoids produced by chemical synthesis. Since carotenoids produced from bacteria have limited industrial applications due to low productivity, studies are continuously being conducted to increase the production of carotenoids by bacteria. Studies conducted to increase carotenoid production from bacteria include the activity of enzymes in the bacterial carotenoid biosynthesis pathway, the development of mutant strains using physical and chemical mutagens, increasing carotenoid productivity in strain construction through genetic engineering, carotenoid accumulation through stress induction, fermentation medium composition, culture conditions, co-culture with other strains, etc. The aim of this article was to review studies conducted to increase the productivity of carotenoids from bacteria.

Short Culm and Early Maturing Mutants Induced by Gamma Irradiation in Rice I . Mutation Rate and Variability (감마선 조사에 의한 수도의 단간 및 조숙돌연변이체 I. 변이체의 출현빈도 및 변이분포)

  • Lee, Young-Il;Shin, In-Chul;Hong, Byeong-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.34 no.4
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    • pp.378-383
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    • 1989
  • For selection of mutants two rice varieties, Sangpungbyeo and Seomjinbyeo, were irradiated with 20 kR and 25 kR of gamma rays. One panicle per each plant was harvested in Ml, and single needling per hill was planted for M2 and M3 generations. Mutations for short culm, earliness and the major traits were examined. 1. The mutation rates were varied with the rice varieties and the doses of radiation, higher in Sangpungbyeo and 25kR than in Seomjinbyeo and 20 kR, respectively. 2. The rates in Sangpungbyeo with the 20 kR and 25 kR were 1.10% and 1.47%. respectively and those of Seomjinbyeo were 0.51% and 1.25%, respectively. 3. The culm lengths of short-culm mutants derived from Sangpungbyeo and Seomjinbyeo were reduced about 10% compared to their mother varieties, especially it was possible to select the dwarf mutants reducing 57% and 40% of culm length in Sangpungbyeo and Seomjinbyeo, respectively with the dose of 25 kR irradiation. 4. The range of heading date of the mutant ion in the M3 generation was comparatively wide. Many earliness mutants shortened about 7 days were selected as compared with their mother varieties, some mutants of the irradiated group had early matured by 20 days and 30 days, respectively in both Seomjinbyeo and Sangpungbyeo.

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Screening for Genotypes Lacking Lipoxygenase from Germplasm Collection of Korean Soybean Land Races (한국 재래종 콩집단에서 비린내 없는 콩품종 육성을 위한 Lipoxygenase 결실인자 변이 연구)

  • Kwon, Shin-Han;Park, Kyung-Sook;Kim, Mi-Young;Kim, Bong-Ryong;Song, Hi-Sup
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.37 no.6
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    • pp.528-533
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    • 1992
  • Soybean seeds contain lipoxygenase, which is responsible for the objectionable beany flavors in soybean seeds. The isozymes of lipoxygenase (1$\times$1, 1$\times$2, 1$\times$3) were discovered in United States of America, Japan, and Korea, and the mode of inheritance of the mutant genes was determined. This investigation was conducted to screen lipoxygenase-1, 2, and 3 lacking soybean lines from the Korean soybean land race population. Two lipoxygenase-1lacking lines, KAS 610-8 and KAS 621-8 were found in this investigation. In general, lipoxygenase acking varieties were small in seed size and low in oil content. A severe pod borer damage was observed in the two selected lipoxygenase-1 lacking lines. Lipoxygenase lacking line was not found in Korean wild soybean population used in this study and the lipoxygenase lacking lines were found only in Kyung-Nam province and the results imply that lipoxygenase lacking mutants were induced recently in cultivars.

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N6-Methyladenosine modification (m6A) of circRNA-ZNF638 contributes to the induced activation of SHF stem cells through miR-361-5p/Wnt5a axis in cashmere goats

  • Ronghuan Yin;Ronglan Yin;Man Bai;Yixing Fan;Zeying Wang;Yubo Zhu;Qi Zhang;Taiyu Hui;Jincheng Shen;Siyu Feng;Wenlin Bai
    • Animal Bioscience
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    • v.36 no.4
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    • pp.555-569
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    • 2023
  • Objective: The objective of this study was to investigate the effects of N6-Methyladenosine modification-circRNA-zinc finger protein 638 (m6A-circRNA-ZNF638) on the induced activation of secondary hair follicle (SHF) stem cells with its potential mechanisms in cashmere goats. Methods: The m6A modification of ZNF638 was analyzed using methylation immunoprecipitation with real-time quantitative polymerase chain reaction technique in SHF stem cells. The effects of circRNA-ZNF638 on the induced activation of SHF stem cells in m6A dependence were evaluated through the overexpression of circRNA-ZNF638/its m6A-deficient mutants in circRNA-ZNF638 knockdown SHF stem cells. The competitive binding of miR-361-5p to circRNA-ZNF638/Wnt5a 3'- untranslated region was analyzed through Dual-luciferase reporter assay. Results: The m6A-circRNA-ZNF638 had significantly higher transcription at anagen SHF bulge of cashmere goats compared with that at telogen, as well as it positively regulated the induced activation of SHF-stem cells in cashmere goats. Mechanismly, m6A-circRNA-ZNF638 sponged miR-361-5p to heighten the transcriptional expression of Wnt5a gene in SHF-stem cells. We further demonstrated that the internal m6A modification within circRNA-ZNF638 is required for mediating the miR-361-5p/Wnt5a pathway to regulate the induced activation of SHF stem cells through an introducing of m6A-deficient mutant of circRNA-ZNF638. Conclusion: The circRNA-ZNF638 contributes the proper induced activation of SHF-stem cells in cashmere goats in m6A-dependent manner through miR-361-5p/Wnt5a axis.

Monitoring trafficking and expression of hemagglutinin-tagged transient receptor potential melastatin 4 channel in mammalian cells

  • Eun Mi Hwang;Bo Hyun Lee;Eun Hye Byun;Soomin Lee;Dawon Kang;Dong Kun Lee;Min Seok Song;Seong-Geun Hong
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.4
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    • pp.417-426
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    • 2023
  • The TRPM4 gene encodes a Ca2+-activated monovalent cation channel called transient receptor potential melastatin 4 (TRPM4) that is expressed in various tissues. Dysregulation or abnormal expression of TRPM4 has been linked to a range of diseases. We introduced the hemagglutinin (HA) tag into the extracellular S6 loop of TRPM4, resulting in an HA-tagged version called TRPM4-HA. This TRPM4-HA was developed to investigate the purification, localization, and function of TRPM4 in different physiological and pathological conditions. TRPM4-HA was successfully expressed in the intact cell membrane and exhibited similar electrophysiological properties, such as the current-voltage relationship, rapid desensitization, and current size, compared to the wild-type TRPM4. The presence of the TRPM4 inhibitor 9-phenanthrol did not affect these properties. Furthermore, a wound-healing assay showed that TRPM4-HA induced cell proliferation and migration, similar to the native TRPM4. Co-expression of protein tyrosine phosphatase, non-receptor type 6 (PTPN6 or SHP1) with TRPM4-HA led to the translocation of TRPM4-HA to the cytosol. To investigate the interaction between PTPN6 and tyrosine residues of TRPM4 in enhancing channel activity, we generated four mutants in which tyrosine (Y) residues were substituted with phenylalanine (F) at the N-terminus of TRPM4. The YF mutants displayed properties and functions similar to TRPM4-HA, except for the Y256F mutant, which showed resistance to 9-phenanthrol, suggesting that Y256 may be involved in the binding site for 9-phenanthrol. Overall, the creation of HA-tagged TRPM4 provides researchers with a valuable tool to study the role of TRPM4 in different conditions and its potential interactions with other proteins, such as PTPN6.

Study of the Effects of Sodium Chloride on Segregation-Distorter Action in D. melanogaster: 1. Theatment of Sodium Chloride on Whole Developmental Stages.

  • Kang, M.J.;Chung, Y.J.;Kim, K.J.
    • The Korean Journal of Zoology
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    • v.13 no.1
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    • pp.21-25
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    • 1970
  • The present paper deals with the results of the experiments, in which the resistibility to NaCl in whole develomental stages is examined by the emergence rate and the effects of NaCl on the SD action in Drosophila melanogaster. The four SD strains and one mutant strain(cn bw)are used and NaCl media are prepared y adding NaCl at a concentration of 1.0M, 0.7M, 0.5M, 0.3M, 0.1M, and 0.0M to the standard media for the present investigations. The results are given below. 1. The emergence rate (resistibility to NaCl) is not significantly different among strains but strikingly different among concentrations of NaCl. 2. The emergence rate decreases as the concentration of NaCl increases; the four SD strains are considerably resistible to the NaCl from a concentration of 0.0M to 0.3M but are susceptible from 0.5M or higher concentrations of NaCl. 3. No eggs are hatched from the culture media containing a concentration of 1.0M NaCl. This suggests that the SD strains are not resistible to NaCl at a concentrations of 1.0M or higher. 4. The difference in k values is not signficant among strains and also among concentrations of NaCl. Thus the SD action is not affected as far as once emerged from the culture media whether containing NaCl or not.

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Global Transcriptome-Wide Association Studies (TWAS) Reveal a Gene Regulation Network of Eating and Cooking Quality Traits in Rice

  • Weiguo Zhao;Qiang He;Kyu-Won Kim;Feifei Xu;Thant Zin Maung;Aueangporn Somsri;Min-Young Yoon;Sang-Beom Lee;Seung-Hyun Kim;Joohyun Lee;Soon-Wook Kwon;Gang-Seob Lee;Bhagwat Nawade;Sang-Ho Chu;Wondo Lee;Yoo-Hyun Cho;Chang-Yong Lee;Ill-Min Chung;Jong-Seong Jeon;Yong-Jin Park
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.207-207
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    • 2022
  • Eating and cooking quality (ECQ) is one of the most complex quantitative traits in rice. The understanding of genetic regulation of transcript expression levels attributing to phenotypic variation in ECQ traits is limited. We integrated whole-genome resequencing, transcriptome, and phenotypic variation data from 84 Japonica accessions to build a transcriptome-wide association study (TWAS) based regulatory network. All ECQ traits showed a large phenotypic variation and significant phenotypic correlations among the traits. TWAS analysis identified a total of 285 transcripts significantly associated with six ECQ traits. Genome-wide mapping of ECQ-associated transcripts revealed 66,905 quantitative expression traits (eQTLs), including 21,747 local eQTLs, and 45,158 trans-eQTLs, regulating the expression of 43 genes. The starch synthesis-related genes (SSRGs), starch synthase IV-1 (SSIV-1), starch branching enzyme 1 (SBE1), granule-bound starch synthase 2 (GBSS2), and ADP-glucose pyrophosphorylase small subunit 2a (OsAGPS2a) were found to have eQTLs regulating the expression of ECQ associated transcripts. Further, in co-expression analysis, 130 genes produced at least one network with 22 master regulators. In addition, we developed CRISPR/Cas9-edited glbl mutant lines that confirmed the role of alpha-globulin (glbl) in starch synthesis to validate the co-expression analysis. This study provided novel insights into the genetic regulation of ECQ traits, and transcripts associated with these traits were discovered that could be used in further rice breeding.

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