• Title/Summary/Keyword: gene-expression

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Transformation of Bottle Gourd Rootstock (Lagenaria siceraria Standl.) using GFP gene (GFP유전자를 이용한 대목용 박 형질전환)

  • Lim, Mi-Young;Park, Sang-Mi;Kwon, Jung-Hee;Han, Sang-Lyul;Shin, Yoon-Sup;Han, Jeung-Sul;Harn, Chee-Hark
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.33-37
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    • 2006
  • Bottle gourd (Lagenaria siceraria Standl.) has been used as a rootstock for the watermelon cultivation because of better growth ability at low temperature and avoidance from contamination of the soil disease. Since the genetic source for the elite rootstock is limited in nature, the genetic engineering method is inevitable to develop new lines especially to obtain the functionally important or multi-disease resistant bottle gourd. Recently, our lab has set up a successful system to transform the bottle gourd. in order to monitor the transformation process, GFP gene is used. Cotyledons of the inbred line 9005, 9006 and G5 were used to induce the shoot under the selection media with MS + 30 g/L sucrose + 3.0 mg/L BAP + 100 mg/L kanamycin + 500 mg/L cefotaxime + 0.5 mg/L $AgNO_3$, pH 5.8. The shoot was developed from the cut side of the explants after 3 weeks on the selection media. The shoot was incubated in the rooting media with 1/2 MS + 30 g/L sucrose + 0.1 mg/L IAA + 50 mg/L kanamycin + 500 mg/L cefotaxime, pH 5.8 and moved to pot for acclimation. Although the shoot development rate was depended on the genotype, the G5 was the best line to be transformed. Monitoring GFP expression from the young shoot under microscope could make the selection much easier to distinguish the transformed shoot from the non-transformed shoots.

Development and Expression of Porcine Embryos by Direct Injection of Sperm Treated with Exogenous DNA (외래유전자 도입정자를 이용한 돼지 체외성숙 난포란의 Intracytoplasmic Sperm Injection (ICSI) 후 후기 배로의 발달율과 외래유전자의 발현에 관한 연구)

  • 정기화;조성근
    • Journal of Embryo Transfer
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    • v.17 no.1
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    • pp.45-53
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    • 2002
  • The main goal of this study was to produce transgenic porcine embryos by direct injection of sperm-mediated exogenous DNA. Spermatozoa (6$\times$10$^{6}$ sperms of final concentration) were mixed with pcDNA LAC Z (20 ng/$\mu$l) and subjected into electroporation (300~750 volts, 25 $\mu$F, 0.4 cm electrode). After sperm injection, the oocytes were activated electrically (1.7 KV/cm, 30$\mu$sec, single pulse) in 0.3 M mannitol solution or not. The sperm injected eggs were cultured in NCSU 23 medium (0.4% BSA) at 39$^{\circ}C$, 5% $CO_2$ in air fur 144 h. The rates of cleavage and development into blastocyst stage in activation group were significantly higher than those of non-activation group (79.6% and 24.1% vs. 46.3% and 14.4%, respectively, p<0.05). Control oocytes and shame injection were developed to blastocysts low (2.5%). Sixty five (27.1%) out of 240 embryos observed in activation and non-activation groups were showed positive by X-gal staining. However, all embryos in both groups were expressed partial or mosaic pattern. These results suggested that electrical stimulation far oocytes activation after sperm injection enhances the incidence of both fertilization and development fellowing sperm injection in the pig. Our study also suggested that sperm-mediated transfer of exogenous DNA by ICSI would be used as a valuable tool for the production of transgenic porcine embryos.

Leaf Shape Index, Growth, and Phytochemicals in Two Leaf Lettuce Cultivars Grown under Monochromatic Light-emitting Diodes (단색 발광다이오드에서 자란 축면상추 두 품종의 엽형, 생장 및 기능성 물질)

  • Son, Ki-Ho;Park, Jun-Hyung;Kim, Daeil;Oh, Myung-Min
    • Horticultural Science & Technology
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    • v.30 no.6
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    • pp.664-672
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    • 2012
  • As an artificial light source, light-emitting diode (LED) with a short wavelength range can be used in closed-type plant production systems. Among various wavelength ranges in visible light, individual light spectra induce distinguishing influences on plant growth and development. In this study, we determined the effects of monochromatic LEDs on leaf shape index, growth and the accumulation of phytochemicals in a red leaf lettuce (Lactuca sativa L. 'Sunmang') and a green leaf lettuce (Lactuca sativa L. 'Grand rapid TBR'). Lettuce seedlings grown under normal growing conditions ($20^{\circ}C$, fluorescent lamp + high pressure sodium lamp, $130{\pm}5{\mu}mmol{\cdot}m^{-2}{\cdot}s^{-1}$, 12 hours photoperiod) for 18 days were transferred into incubators at $20^{\circ}C$ equipped with various monochromatic LEDs (blue LED, 456 nm; green LED, 518 nm; red LED, 654 nm; white LED, 456 nm + 558 nm) under the same light intensity and photoperiod ($130{\pm}7{\mu}mmol{\cdot}m^{-2}{\cdot}s^{-1}$, 12 hours photoperiod). Leaf length, leaf width, leaf area, fresh and dry weights of shoots and roots, shoot/root ratio, SPAD value, total phenolic concentration, antioxidant capacity, and the expression of a key gene involved in the biosynthesis of phenolic compounds, phenylalanine ammonia-lyase (PAL), were measured at 9 and 23 days after transplanting. The leaf shape indexes of both lettuce cultivars subjected to blue or white LEDs were similar with those of control during whole growth stage. However, red and green LEDs induced significantly higher leaf shape index than the other treatments. The green LED had a negative impact on the lettuce growth. Most of growth characteristics such as fresh and dry weights of shoots and leaf area were the highest in both cultivars subjected to red LED treatment. In case of red leaf lettuce plants, shoot fresh weight under red LED was 3.8 times higher than that under green LED at 23 days after transplanting. In contrast, the accumulation of chlorophyll, phenolics including antioxidants in lettuce plants showed an opposite trend compared with growth. SPAD value, total phenolic concentration, and antioxidant capacity of lettuce grown under blue LED were significantly higher than those under other LED treatments. In addition, PAL gene was remarkably activated by blue LED at 9 days after transplanting. Thus, this study suggested that the light quality using LEDs is a crucial factor for morphology, growth, and phytochemicals of two lettuce cultivars.

Isolation and characterization of sigH from Corynebacterium glutamicum (Corynebacterium glutamicum의 sigH 유전자의 분리 및 기능분석)

  • Kim Tae-Hyun;Kim Hyung-Joon;Park Joon-Sung;Kim Younhee;Lee Heung-Shick
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.99-104
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    • 2005
  • Corynebacterial clones which exert regulatory effects on the expression of the glyoxylate bypass genes were isolated using a reporter plasmid carrying the enteric lacZ fused to the aceB promoter of Corynebacterium glutamicum. Some clones carried common fragments as turned out by DNA mapping technique. Subcloning analysis followed by the measurement of $\beta-galactosidase$ activity in Escherichia coli identified the region responsible for the aceB-repressing activity. Sequence analysis of the DNA fragment identified two independent ORFs of ORF1 and ORF2. Among them, ORF2 was turned out to be responsible for the aceB-repressing activity. ORF1 encoded a 23,216 Da protein composed of 206 amino acids. Sequence similarity search indicated that the ORF may encode a ECF-type $\sigma$ factor and designated sigH. To identify the function of sigH, C. glutamicum sigH mutant was constructed by gene disruption technique and the sigH mutant showed growth retardation as compared to the wild type strain. In addition, the mutant strain showed sensitivity to oxidative-stress generating agent plumbagin. This result imply that sigH is probably involved in the stress response occurring during normal cell growth.

The Functional and Genetic Defects of IFN-${\gamma}$ Receptor in the Patients with Tuberculosis (결핵환자에서 IFN-${\gamma}$ 수용체의 기능적 및 유전적 이상에 관한 연구)

  • Park, Gye-Young;Hwang, You-Jin;Lim, Young-Hee;An, Chang-Hyeok;Park, Jeong-Woong;Jeong, Seong-Hwan
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.5
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    • pp.497-505
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    • 2002
  • Background : INF-${\gamma}$ plays an important role in the host response to a mycobacterial infection. A complete IFN-${\gamma}$ receptor 1 deficiency is a life threatening condition because it renders patients highly susceptible to a mycobacterial infection. Several mutations in the IFN-${\gamma}$ receptor and STAT1 gene have been identified in the rare mycobacterial infections. These mutations have partial function of the IFN-${\gamma}$ receptor and similar pathologic features to clinical tuberculosis. Materials and Methods : The function of the IFN-${\gamma}$ receptor was evaluated in the patients with clinical tuberculosis. In addition, the DNA coding sequence of the IFNgR1 and STAT1 gene was also analyzed in disseminated tuberculosis patients who might have a defective IFN-${\gamma}$ receptor. Results : The cell surface expression levels of HLA-DR and CD64 in the PMBC after being stimulation with IFN-${\gamma}$ (100IU/ml, 1000IU/ml) were increased in both controls and patients. However, the rate of increase in both groups was similar. The production of TNF-${\alpha}$ in the response to stimulation with LPS was higher in the both groups ($850.7{\pm}687.8$ vs. $836.7{\pm}564.3$ pg/ml). Pretreatment with IFN-${\gamma}$ prior to LPS stimulation resulted in further increase in TNF-${\alpha}$ production between both groups ($2203.5{\pm}242.5$ vs. $2227.5{\pm}560.4$ pg/ml). However, the rate of the increase in TNF-${\alpha}$ production in the both groups was similar. The known mutations in the IFNgR1 and STAT1 coding sequences were not found in the genomic DNA of patients with disseminated tuberculosis. Conclusion : The functional and genetic defects of the IFN-${\gamma}$ receptor were not identified in clinical tuberculosis. This suggests the defective IFN-${\gamma}$ receptor that predispoe patients to a BCG or NTM infection can not alone account for the cases of clinical tuberculosis.

Antioxidative Activities of the Codonopsis lanceolata Extract in vitro and in vivo (더덕(Codonopsis lanceolata) 추출물의 in vitro 및 in vivo 항산화 효과)

  • Kim, Soo-Hyun;Chung, Mi-Ja;Jang, Hae-Dong;Ham, Seung-Shi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.2
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    • pp.193-202
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    • 2010
  • In vitro activities of Codonopsis lanceolata (CL) 70% ethanol extract and its fractions (hexane, chloroform, ethyl acetate, butanol and water) were examined by total polyphenol content, reducing power, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS), 2,2-diphenyl-$\beta$-picrylhydrazyl (DPPH), and oxygen radical absorbance capacity (ORAC) assays. The ethyl acetate fraction from CL ethanol extract (CLEA) showed the highest total polyphenol content (22.7 mg/g) among five fractions, and also exhibited an excellent reducing power (0.42~1.27 at $250\sim1,000\;{\mu}g/mL$). CLEA at $100\sim400\;{\mu}g/mL$ concentrations had 27.7~70.3% of ABTS radical scavenging activity and the highest DPPH radical scavenging activity (81.6% at $400\;{\mu}g/mL$). CLEA had dominantly higher $ORAC_{{ROO}{\cdot}}$activity compared to other fractions. CLEA and butanol fraction had significantly higher $ORAC_{{OH}{\cdot}}$ activities than 70% ethanol extract, hexane, chloroform and water fractions. The CLEA exhibited the highest antioxidant activity in CL 70% ethanol extract and its fractions. Thus, effect of CLEA treatment on antioxidant gene expression under the oxidative stress conditions by a high fat diet in animal model was studied by microarray and RT-PCR methods. The 31 antioxidant genes were expressed but the genes were not up-regulated at least a two-fold by CLEA treatment. We concluded that CLEA does not have an indirect antioxidant effect but a direct antioxidant effect by up-regulation of antioxidant genes in high fat diet-induced obese mice.

The Effect of Exercise Training on Aβ-42, BDNF, GLUT-1 and HSP-70 Proteins in a NSE/ APPsw-transgenic Model for Alzheimer's Disease. (지구성 운동이 NSE/APPsw 알츠하이머 질환 생쥐의 인지능력, Aβ-42, BDNF, GLUT-1과 HSP-70 단백질 발현에 미치는 영향)

  • Eum, Hyun-Sub;Kang, Eun-Bum;Lim, Yea-Hyun;Lee, Jong-Rok;Cho, In-Ho;Kim, Young-Soo;Chae, Kab-Ryoung;Hwang, Dae-Yean;Kwak, Yi-Sub;Oh, Yoo-Sung;Cho, Joon-Yong
    • Journal of Life Science
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    • v.18 no.6
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    • pp.796-803
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    • 2008
  • Mutations in the APP gene lead to enhanced cleavage by ${\beta}-$ and ${\gamma}-secretase$, and increased $A{\beta}$ formation, which are closely associated with Alzheimer's disease (AD)-like neuropathological changes. Recent studies have shown that exercise training can ameliorate pathogenic phenotypes ($A{\beta}-42$, BDNF, GLUT-1 and HSP70) in experimental models of Alzheimer's disease. Here, we have used NSE/APPsw transgenic mice to investigate directly whether exercise training ameliorates pathogenic phenotypes within Alzheimer's brains. Sixteen weeks of exercise training resulted in a reduction of $A{\beta}-42$ peptides and also facilitated improvement of cognitive function. Furthermore, GLUT -1 and BDNF proteins produced by exercise training may protect brain neurons by inducing the concomitant expression of genes that encode proteins (HSP-70) which suppress stress induced neuron cell damages from APPsw transgenic mice. Thus, the improved cognitive function by exercise training may be mechanistically linked to a reduction of $A{\beta}-42$ peptides, possibly via activation of BDNF, GLUT-1, and HSP-70 proteins. On the basis of the evidences presented in this study, exercise training may represent a practical therapeutic management strategy for human subjects suffering from Alzheimer's disease.

FT-transgenic spray-type Chrysanthemum (Dendranthema grandiflorum Kitamura) showing early-flowering (FT 유전자 형질전환 스프레이 국화 (Dendranthema grandiflorum (Ramat.) Kitamura)의 조기개화성)

  • Lee, Su-Young;Han, Bong-Hee;Hur, Eun-Joo;Shin, Hak-Kee;Lee, Il-Ha;Lee, Eun-Kyung;Kim, Seung-Tae;Kim, Won-Hee;Kwon, O-Hyeon
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.140-145
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    • 2012
  • The flowering locus T (FT) gene, of which expression will be controlled at high temperature by heat shock promoter (it printed as to HSproFT), was introduced into spray-type chrysanthemum (Dendranthema grandiflorum (Ramat.) Kitamura) 2 cultivars ('Pink PangPang' and 'Pink Pride' by co-cultivation with Agrobacterium tumefaciens strain C58C1 harboring pCAMBIA2300 containing the HSproFT gene. After leaf segments of the 2 cultivars were infected with the A. tumafaciens with C58C1 as explants, shoots were regenerated from the explants cultured on the $1^{st}$ selection medium (MS basal salts + 1.0 mg/L BA, 0.5 mg/L IAA + 10 mg/L kanamycin + 0.7% plant agar, pH 5.8). The shoots were transferred into the $2^{nd}$ selection medium (MS basal salts + 1.0 mg/L BA, 0.5 mg/L IAA + 20 mg/L kanamycin + 0.7% plant agar, pH 5.8). One hundred seventeen plantlets from 'Pink PangPang' and 5 ones from 'Pink Pride' were confirmed as transformants by PCR analysis. Twenty six of the transformants and non-transformants were acclimatized and established well in a green house. Eights of 26 transgenic lines showed flower bud 1.7~10 days earlier than nontransgenic plants, and 24 of them flowered 1~6 days earlier than non- transgenic plants. The shape and color of flower of all HSproFT-transgenic lines were not different with those of non- transgenic plants.

Estrogeicity of Genistein and Bisphenol A (콩류식품의 주성분인 Genistein과 식품포장재 및 용기에 사용되는 Bisphenol A의 에스트로젠 효과에 관한 연구)

  • 강경선;이영순;신광순
    • Journal of Food Hygiene and Safety
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    • v.13 no.2
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    • pp.106-111
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    • 1998
  • This study has been focused on both estrogenic and proliferating activity of genistein (GEN) and bisphenol A (BPA). GEN and BPA enhance the proliferation of estrogen-dependent MCF-7 human breast cancer cells at concentrations as low as 100 nM of GEN and 8 ng/ml of BP A achieving similar effect to that of estradiol at 1 nM. Expression of the estrogen responsive gene, pS2 was also induced in MCF-7 cells by treatment with genistein at dose as low as 1 nM and BPA at dose as low as 4 ng/ml. Using 21 day-old ovariectomized nude mice, we examined end-bud formation and mammary gland development after treatment with bisphenol A or genistein. Compared with untreated control, mammary gland development and end-bud formation were significantly increased in mice fed genistein or bisphenol A (p<0.05). Taken together, it is concluded that GEN and BP A can act as an estrogen agonist resulting in cell proliferation and induction of the estrogen responsive pS2 gene in MCF-7 cells in vitro and in athymic mice in vivo, respectively. Therefore, it is suggested that GEN and BP A might modulate human endocrine system and these compounds might be considered as a endocrine modulator at the low levels of doses.

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Ethanol Extracts of Mori Folium Inhibit Adipogenesis Through Activation of AMPK Signaling Pathway in 3T3-L1 Preadipocytes (3T3-L1 세포에서 상엽이 유발하는 AMPK signaling pathway를 통한 adipogenesis 억제에 관한 연구)

  • Ji, Seon Young;Jeon, Keong Yoon;Jeong, Jin Woo;Hong, Su Hyun;Huh, Man Kyu;Choi, Yung Hyun;Park, Cheol
    • Journal of Life Science
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    • v.27 no.2
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    • pp.155-163
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    • 2017
  • Mori Folium, the leaf of Morus alba, is a traditional medicinal herb that shows various pharmacological activities such as antiinflammatory, antidiabetic, antimelanogenesis, antioxidant, antibacterial, antiallergic, and immunomodulatory activities. However, the mechanisms of their inhibitory effects on adipocyte differentiation and adipogenesis remain poorly understood. In the present study, we investigated the inhibition of adipocyte differentiation and adipogenesis by ethanol extracts of Mori Folium (EEMF) in 3T3-L1 preadipocytes. Treatment with EEMF suppressed the terminal differentiation of 3T3-L1 preadipocytes in a dose-dependent manner, as confirmed by a decrease in the lipid droplet number and lipid content through Oil Red O staining. EEMF significantly reduced the accumulation of cellular triglyceride, which is associated with a significant inhibition of pro-adipogenic transcription factors, including sterol regulatory element-binding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor-${\gamma}$ ($PPAR{\gamma}$), and CCAAT/enhancer-binding proteins ${\alpha}$ ($C/EBP{\alpha}$) and ${\beta}$ ($C/EBP{\beta}$). In addition, EEMF potentially downregulated the expression of adipocyte-specific genes, including adipocyte fatty acid binding protein (aP2) and leptin. Furthermore, EEMF treatment effectively increased the phosphorylation of the AMP-activated protein kinase (AMPK) and acetyl CoA carboxylase (ACC); however, treatment with a potent inhibitor of AMPK, compound C, significantly restored the EEMF-induced inhibition of pro-adipogenic transcription factors and adipocyte-specific genes. These results together indicate that EEMF has preeminent effects on the inhibition of adipogenesis through the AMPK signaling pathway, and further studies will be needed to identify the active compounds in Mori Folium.