• Title/Summary/Keyword: nuclear DNA content

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Construction of Starch-Fermenting Yeast Using Protoplast Fusion Technique (원형질체 융합 방법을 이용한 전분 발효성 효모의 개발)

  • Chun, Soon-Bai;Bai, Suk;Lee, Kee-Young;Kim, Soo-Hyun;Lee, Jin-Jong;Kim, Sang-Moon
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.120-127
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    • 1990
  • Intraspecific fusion frequency between Filobasidium capsuligenum CBS6122-2 ade and met trp or met strains was $2.9\times 10^{-3}$ and $8.5\times 10^{-3}$, respectively. And intergeneric fusion frequency between Folobadidium capsuligenum CBS 4318 (cys his) and Saccharomyces cerevisiae 262-12-1 (hom3 thr1) or X2180-1A (ade thr) was $8.8\times 10^{-6}$ and $9.5\times 10^{-4}$, respectively. Nuclear fusion appears to occru in fusants between intraspecies and intergenera as strongly suggested by DNA content, nuclear staining, comparison of survival rate to UV light and mitotic segregation analysis. It was also found that $\alpha$-amylase and glucoamylase activity from intraspecific hybrids was 1.6-2.1 fold increased when compared with that from thier parents.

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Genetic Diversity among Korean Bermudagrass (Cynodon spp.) Ecotypes Characterized by Morphological, Cytological and Molecular Approaches

  • Kang, Si-Yong;Lee, Geung-Joo;Lim, Ki Byung;Lee, Hye Jung;Park, In Sook;Chung, Sung Jin;Kim, Jin-Baek;Kim, Dong Sub;Rhee, Hye Kyung
    • Molecules and Cells
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    • v.25 no.2
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    • pp.163-171
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    • 2008
  • The genus Cynodon comprises ten species. The objective of this study was to evaluate the genetic diversity of Korean bermudagrasses at the morphological, cytological and molecular levels. Morphological parameters, the nuclear DNA content and ploidy levels were observed in 43 bermudagrass ecotypes. AFLP markers were evaluated to define the genetic diversity, and chromosome counts were made to confirm the inferred cytotypes. Nuclear DNA contents were in the ranges 1.42-1.56, 1.94-2.19, 2.54, and 2.77-2.85 pg/2C for the triploid, tetraploid, pentaploid, and hexaploid accessions, respectively. The inferred cytotypes were triploid (2n = 3x = 27), tetraploid (2n = 4x = 36), pentaploid (2n = 5x = 45), and hexaploid (2n = 6x = 54), but the majority of the collections were tetraploid (81%). Mitotic chromosome counts verified the corresponding ploidy levels. The fast growing fine-textured ecotypes had lower ploidy levels, while the pentaploids and hexaploids were coarse types. The genetic similarity ranged from 0.42 to 0.94 with an average of 0.64. UPGMA cluster analysis and principle coordinate analysis separated the ecotypes into 6 distinct groups. The genetic similarity suggests natural hybridization between the different cytotypes, which could be useful resources for future breeding and genetic studies.

Cytogenetic Analysis of Spotty Belly Greenling (Hexagrammos agrammus) and Greenling (H. otakii) (노래미 (Hexagrammos agrammus)와 쥐노래미 (H. otakii)의 세포유전학적 연구)

  • Sim Mi A;Noh Jae Koo;Nam Yoon Kwon;Kim Dong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.6
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    • pp.682-685
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    • 2002
  • Cytogenetic analysis was conducted to obtaining informations for genetic improvement of spotty belly greenling (Hexagrmmos agrammus) and greenling (H. otakii) in aquaculture. Erythrocytes of spotty belly greenling were slightly larger than those of greenling (p<0.05). The nuclear volume of spotty belly greening erythrocytes averaged 15.14 $\pm$ 0.92 ${\mu}m^3$ while that of greening averaged 14.61 $\pm$ 0.15 $\mu$m^3 the difference was not significant (p>0.05). Consequently, genome size of spotty belly greenling was also slightly larger than those of greenling. DNA content per cell of spotty belly greenling and greenling were 2.15 pg and 2.10 pg, respectively. The modal chromosome number of both greenling species were same as 2n=48 and karyotypes were also identical as 2 metacentrics, 11 snbrnetacentrics and 11 acrocentric pairs $(W: 74), There was no evidence of polymorphism including aneuploidy or sex-related heterornorphisrn for all specimens examined. The nuclear organizer regions (NOR_s)$ were localized on a small acrocentric chromosome pair in both species, Spotty belly greenling showed large sizes of active rRNA coding regions in their chromosomes. However, greenling examined only small sizes of active rRNA coding regions with dimorphism.

Formation of Intergeneric Hybrids Between Aspergillus niger and Penicillium verruculosum by Nuclear Transfer (핵전이에 의한 Aspergillus niger와 Penicillium verruculosum F-3의 속간 잡종형성)

  • Yang, Young-Ki;Park, Yeol;Kim, Sung-Joon;Cheong, Hyeon-Suk;Lim, Chae-Young;Rhee, Young-Ha
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.1-8
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    • 1993
  • lntergeneric hybrids formed between Aspergillus niger and Penicillium verruculosum were obtained by nuclear transfer technique. Nuclei isolated from wild type and auxotrophic mutants of donor strains were transferred into the protoplasts of different auxotrophic mutants as recipient strains. Several auxotrophic mutants were isolated from conidiospores of the two strains mutagenized with ultraviolet and N-methyl-N'-nitrosoguanidine. Optimal conditions for formation of intergeneric hybrids were investigated. Frequencies of intergeneric hybrid formation by nuclear transfer were $7{\times}10^{5}~1{\times}10^{5}$. From observations of genetic stability. DNA content. nuclear stain and conidial size. it was suggested that their karyotypes are aneuploid. In addition. the hybrids possess the 1.1~2.3-fold higher cellulase activities than those of parental strains. It was also revealed that some hybrids had different isozyme patterns compared to those of parental strains by CMCase and $\beta$-glucosidase activity assays.

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Cytogenetic Analysis of Starry Flounder Platichthys stellatus from Korea (강도다리(Platichthys stellatus)에 대한 세포유전학적 연구)

  • Jung, Hyo Sun;Kim, Youn Kyoung;Kim, Hyun Chul;Noh, Jae-Koo;Lee, Jong-Ho;Kim, Dong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.47 no.4
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    • pp.431-434
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    • 2014
  • Cytogenetic analysis was conducted to obtain basic information for chromosome manipulation of starry flounder Platichthys stellatus. Nuclear surface area and volume of erythrocyte were $7.60{\pm}0.93{\mu}m^2$ and $12.80{\pm}1.75{\mu}m^3$, respectively. The haploid DNA content of the species was 0.66 pg/haploid cell which correspond to 93% of olive flounder Paralichthys olivaceus. A karyotype analysis was also carried out with the species using conventional staining and Ag-NOR banding techniques. It was consisted of 48 acrocentric chromosomes and inter-sex or intra-individual polymorphism was not detected in all specimens analyzed. The NOR regions, appearing a terminal position of the short arm of the smallest acrocentric pairs.

Cytogenetic Analysis of Induced Hybrid between Common Carp (Cyprinus carpio) and Crucian Carp (Carassius auratus) (잉어(Cyprinus carpio)와 붕어(Carassius auratus)간 잡종의 세포유전학적 분석)

  • 남윤권;오승용;조재윤;김동수
    • Journal of Aquaculture
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    • v.11 no.1
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    • pp.77-81
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    • 1998
  • Cytogenetic analyses were carried out with induced hybrid between common carp (Cyprinus carpio) female and crucian carp (Carassius auratus) male. The erythrocytic measurement revealed that cellular and nuclear size of induced hybrids were intermediate between those of paremtal species. The modal chromosome numbers of common carp, crucian carp and its hybid were same as 2n=100. The DNA content of induced hybrids determined based on flow cytometry was 3.7pg/cell which corresponding to intermediate value between the carp (3.6ph/cell) and crucian carp (3.8pg/cell)

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Long-Term Effects of Growth Regulators and Nitrogen Sources on Proliferation and Turnover of Cell Wall Polysaccharides in Suspension Culture of Kidney Bean (Phaseolus vulgaris L.) (강낭콩의 현탁배양시 증식과 세포벽 다당류 전환에 미치는 생장조절제 및 질소원의 장기간 효과)

  • CHAI, Youn Kyung;KIM, Kyong Ho;YEO, Up Dong;SAKURAI Naoki
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.6
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    • pp.477-485
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    • 1998
  • To underatand in vitro regulation of differentiation, the effects of growth regulators and nitrogen source on metabolism of cell wall polysaccharides in suspension culture of kidney bean (Phaseolus vulgaris L.) were investigated. The suspension cells (cell clusters) were directly induced from the epicotyl segments of the seedlings, which were cultivated in MS medium supplemented with 1.0mg/L of 2,4-D and 0.5 mg/L of kinetin. When compared with cell wall sugar contents of the epicotyl segments, the cellulose content of the suspension-cultured cells decreased; while the pectin and hemicellulose content increased; suggesting increases of rhamnogalacturonan I and arabinogalactan IIduring the dedifferentiation, respectively, The effects of growth regulators(2,4-D, 1.0mg/L and kinetin, 0.5mg/L) and nitrogen source (potasium nitrate, 19.0mg/L and ammonium nitrate, 16.5 g/L) in the medium on the proliferation and the turnover of the cell wall polysaccharides were investigated for 30 days. In the medium with growth regulators and without nitrogen source, the proliferation rate was extremely high (16 folds). Growth regulators and nitrogen source increased the pectin content. Analysis of neutral sugar composition of pectin fraction showed that nitrogen source enhanced rhamnose level remarkably, suggesting that rhamnogalacturonan I was the one most likely synthesized. In hemicellulose fraction, growth regulators reduced arabinose level, suggesting that arabinogalactan II was degraded. And nitrogen source reduced galactose level, suggesting that xyloglucan was also degraded.

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Effects of dietary leucine supplementation on the hepatic mitochondrial biogenesis and energy metabolism in normal birth weight and intrauterine growth-retarded weanling piglets

  • Su, Weipeng;Xu, Wen;Zhang, Hao;Ying, Zhixiong;Zhou, Le;Zhang, Lili;Wang, Tian
    • Nutrition Research and Practice
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    • v.11 no.2
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    • pp.121-129
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    • 2017
  • BACKGROUND/OBJECTIVES: The study was conducted to evaluate the effects of dietary leucine supplementation on mitochondrial biogenesis and energy metabolism in the liver of normal birth weight (NBW) and intrauterine growth-retarded (IUGR) weanling piglets. MATERIALS/METHODS: A total of sixteen pairs of NBW and IUGR piglets from sixteen sows were selected according to their birth weight. At postnatal day 14, all piglets were weaned and fed either a control diet or a leucine-supplemented diet for 21 d. Thereafter, a $2{\times}2$ factorial experimental design was used. Each treatment consisted of eight replications with one piglet per replication. RESULTS: Compared with NBW piglets, IUGR piglets had a decreased (P < 0.05) hepatic adenosine triphosphate (ATP) content. Also, IUGR piglets exhibited reductions (P < 0.05) in the activities of hepatic mitochondrial pyruvate dehydrogenase (PDH), citrate synthase (CS), ${\alpha}$-ketoglutarate dehydrogenase (${\alpha}$-KGDH), malate dehydrogenase (MDH), and complexes I and V, along with decreases (P < 0.05) in the concentration of mitochondrial DNA (mtDNA) and the protein expression of hepatic peroxisome proliferator-activated receptor-${\gamma}$ coactivator $1{\alpha}$ (PGC-$1{\alpha}$). Dietary leucine supplementation increased (P < 0.05) the content of ATP, and the activities of CS, ${\alpha}$-KGDH, MDH, and complex V in the liver of piglets. Furthermore, compared to those fed a control diet, piglets given a leucine-supplemented diet exhibited increases (P < 0.05) in the mtDNA content and in the mRNA expressions of sirtuin 1, PGC-$1{\alpha}$, nuclear respiratory factor 1, mitochondrial transcription factor A, and ATP synthase, $H^+$ transporting, mitochondrial F1 complex, ${\beta}$ polypeptide in liver. CONCLUSIONS: Dietary leucine supplementation may exert beneficial effects on mitochondrial biogenesis and energy metabolism in NBW and IUGR weanling piglets.

Fusion Hybrid and Carotenoid Formation from the Yeast, Phaffia rhodozyma (효모 Phaffia rhodozyma의 융합체와 Carotenoid 생성)

  • Chang, Kee-Myung;Kim, Moon-Hee;Song, Myung-Hee;Kim, Sang-Moon;Chun, Soon-Bai
    • The Korean Journal of Mycology
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    • v.21 no.1
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    • pp.9-15
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    • 1993
  • ABSTRACT: The cell fusants were constructed from complementary auxotrophic mutants of Phffia rhodozyma. The nuclear fusion of the fusants was demonstrated by several techniques including comparision of cell volume, estimation of DNA content and nuclear staining. The cell fusants were very stable for succeeding transfer culture on complex medium for more than one year. Malt extract (1%, w/v) and abscisic acid(1 mM) increased the carotenoid formation whereas gibberellic acid(5mM) and riboflavin(0.1 mM) decreased the corresponding content.

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Mechanism Underlying NaF-Induced Apoptosis in Human Oral Squamous Cell Carcinoma

  • Hur, Young-Joo;Kim, Do-Kyun;Lee, Seung-Eun;Kim, In-Ryoung;Jeong, Na-Young;Kim, Ji-Young;Park, Bong-Soo
    • International Journal of Oral Biology
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    • v.35 no.2
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    • pp.51-60
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    • 2010
  • Few studies have evaluated the apoptosis-inducing efficacy of NaF on cancer cells in vitro but there has been no previous investigation of the apoptotic effects of NaF on human oral squamous cell carcinoma cells. In this study, we have investigated the mechanisms underlying the apoptotic response to NaF treatment in the YD9 human squamous cell carcinoma cell line. The viability of YD9 cells and their growth inhibition were assessed by MTT and clonogenic assays, respectively. Hoechst staining, DNA electrophoresis and TUNEL staining were conducted to detect apoptosis. YD9 cells were treated with NaF, and western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, and MMP and proteasome activity assays were performed sequentially. The NaF treatment resulted in a time- and dose-dependent decrease in YD9 cell viability, a dose-dependent inhibition of cell growth, and the induction of apoptotic cell death. The apoptotic response of these cells was manifested by nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, a decreased DNA content, the release of cytochrome c into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, a significant shift of the Bax/Bcl-2 ratio, and the activation of caspase-9, caspase-3, PARP, Lamin A/C and DFF45 (ICAD). Furthermore, NaF treatment resulted in the downregulation of G1 cell cyclerelated proteins, and upregulation of p53 and the Cdk inhibitor $p27^{KIP1}$. Taken collectively, our present findings demonstrate that NaF strongly inhibits YD9 cell proliferation by modulating the expression of G1 cell cycle-related proteins and inducing apoptosis via mitochondrial and caspase pathways.