• Title/Summary/Keyword: 유전자독성

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Synergistic Cytotoxic Effects by Combination Treatment of Genistein and Daidzein in Human Colorectal Cancer Cell (대장암 세포주에서 genistein과 daidzein의 병합처리에 의한 상승적인 세포독성 효과)

  • Son, Seong-Min;Lim, Seung-Hyun;Kim, Hyo-Rim;Kim, Min-Jeong;Kim, Tae-Wan;Lee, Jong-Hwa;Kim, Jong-Sik
    • Journal of Life Science
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    • v.19 no.9
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    • pp.1294-1298
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    • 2009
  • To investigate whether isoflavone genistein and daidzein could affect cancer cell viability, human colorectal HCT116 cells were incubated with genistein or daidzein in a dose-dependent manner. Genistein decreased cancer cell viability in a dose-dependent manner, whereas daidzein did not show dramatic cytotoxic effects. We also found that 71 genes were up-regulated more than 2-fold, whereas 64 genes were down-regulated more than 2-fold with 24 hr of $50{\mu}M$ genistein treatment by our previous microarray data. Among the up-regulated genes, we selected 3 genes (DKK1, ATF3 and NAG-1) and performed RT-PCR to confirm microarray data. The results of RT-PCR were highly correlated with those of the microarray experiment. In addition, we investigated whether a combination treatment of genistein and daidzein could affect cancer cell viability. Surprisingly, the combination treatment did show synergistic cytotoxic effects detected by MTS assay. The results of RT-PCR and real-time PCR indicate that a combination of genistein and daidzein can synergistically induce NAG-1 expression in HCT116 cells. This result implies that NAG-1 induction is highly associated with synergistic cytotoxic effects induced by a combination treatment of genistein and daidzein. Overall, these results may provide a clue in explaining the anti-cancer activity of soy bean in human colorectal cancer.

Cloning and Expression of an Insecticidal Crystal Protein CryIIA Gene from Bacillus thuringiensis subsp. kurstaki HD-1 (Bacillus thuringiensis subsp. kurstaki HD-1 CryIIA의 내독소 단백질 유전자의 클로닝 및 발현)

  • 김호산;김상현;제연호;유용만;서숙재;강석권;조용섭
    • Korean journal of applied entomology
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    • v.32 no.3
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    • pp.300-306
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    • 1993
  • The CryIIA gene encoding the insecticidal crystal protein of Bacillus thuringiens!s subsp. kurstalri HD-l has been cloned in Escherichia col!, and its nucleotide sequences were determined completely. 5kb Hindlli fragment harboring CryIIA gene was screened in the large ca. 225kb plasmid DNA by southern blot. HindlIT digested 5kb fragment was ligated into pUC19 and transformed in E. coli. The 4kb BamHI-HindlIT fragment containing the CryIIA gene was subcloned and named pSKIIA. DNA sequence analysis demonstrates that pSKIIA is the gene of an operon which is comprised of Lhree open reading frames (designated orn, orf2 and or£3). The CrylIA gene is composed of 3,952bp-long BamHI-Hindill DNA restriction fragment. The orf3 code for a polypeptide of 633 amino acid residues. The protoxin protein has a predicted molecular weight of 70,780. The E. coli derived protoxin gene product is biologICally active against three species of Lepidopteran (Plu.lelia maculipennis, He/iolhis assulta, Spodoptera litura) and a species of Dip Leran( Culex pipines) larvae in bioassay.

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Regulation of Unfolded Protein Response by Ethylene Glycol in Rat (Rat에서 ethylene glycol에 unfolded protein response의 조절)

  • Lee, Eun Ryeong;Kwon, Kisang
    • Journal of Life Science
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    • v.23 no.9
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    • pp.1104-1108
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    • 2013
  • Ethylene glycol (EG) is the most commonly used for automotive antifreeze, and it's easily misuseful for human. EG poisoning occurs in suicide attempts and infrequently, either intentionally through misuse or accidentally because of sweet taste. Though EG itself is mild toxic to the human body, it becomes higher toxic organic acids by in vivo broken down that are responsible for extensive cellular damage in various tissues caused principally by the metabolites. It is already well known that various cellular stresses induce gene expression of endoplasmic reticulum (ER) chaperones and ER stress sensors. This study demonstrated that regulation of gene expression of ER chaperones and ER stress sensors was induced by EG in rat tissues, and in tissues histological changes are also detected by both staining H&E and immunofluorescent.

Effect of Cryptochlorogenic Acid Extracted from Fruits of Sorbus commixta on Osteoblast Differentiation (마가목 열매에서 추출한 Cryptochlorogenic Acid 처리에 의한 조골세포 분화 촉진 효능)

  • Kim, Kyeong-Min;Kim, Tae Hoon;Jang, Won-Gu
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.3
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    • pp.314-319
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    • 2017
  • Chlorogenic acid, a well-known polyphenol, and its derivatives, ester of caffeic acid on quinic acid moiety, are abundant in coffee, tea, fruits, and various vegetables. This study examined the effects of cryptochlorogenic acid (CCA) on osteoblast differentiation. CCA-induced mRNA expression levels of osteogenic genes in MC3T3E1 and C3H10T1/2 cells were determined by RT-PCR and qPCR. CCA regulated expression of key osteogenic genes in the early stage of differentiation, including distal-less homeobox 5 (Dlx5), DNA-binding protein inhibitor (Id1), and runt-related transcription factor 2 (Runx2). These results suggest that CCA may enhance osteoblast differentiation through expression of osteogenic genes such as Id1, Dlx5, and Runx2, especially in the early stage.

Purification and risk assessment of Bacillus thuringiensis Vip3Aa protein against Apis mellifera (Bacillus thuringiensis 유래 Vip3Aa 단백질 순수분리 및 꿀벌 (Apis mellifera)에 대한 위해성평가)

  • Jung, Young Jun;Yoo, Su-Hyang;Lee, Jung Ro
    • Korean Journal of Environmental Biology
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    • v.37 no.4
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    • pp.585-591
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    • 2019
  • Most insect-resistant LMOs have been produced by applying Cry and Vip3Aa proteins. Vip3Aa protein is activated during the vegetative stage of Bacillus thuringensis (Bt) and the inhibitory activity of the Vip3Aa protein against pathogenic attacks from lepidopteran insect species is well known. However, a risk assessment of the Vip3Aa protein compared to the Cry protein has not been conducted in South Korea. This study demonstrates a possible risk assessment method for Vip3Aa protein against honeybees (Apis mellifera). For the risk assessment of the protein, we purified the recombinant Vip3Aa protein in Escherichia coli. The survival rate and symptoms of general intoxication of 4 months honeybees were measured after Vip3Aa exposure. These results indicated that there was no significant difference in the survival rate and the symptom between Vip3Aa and the control buffer. In this study, we established standard methods of Vip3Aa protein purification and oral adult toxicity test using A. mellifera as an LMO risk assessment technique for preserving the natural ecosystem of South Korea.

Distribution of Toxic and Non-toxic Microcystis in Korean Water Supply (국내 주요 상수원지에서 독성 및 비독성 Microcystis의 분포 특성)

  • Lee, Kyung-Lak;Shin, Yuna;Lee, Jaean;Lee, Jae-Kwan;Kim, Han Soon
    • Korean Journal of Ecology and Environment
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    • v.49 no.4
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    • pp.393-399
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    • 2016
  • This study was conducted to investigate whether the presence of mcy gene and microcystin production are related to morphological characteristics of Korean Microcystis species. We isolated 6 different species of Microcystis (M. aeruginosa, M. ichthyoblabe, M. flos-aquae, M. novacekii, M. viridis and M. wesenbergii) from drinking water supply dams (Yeongchun, Ankei, Gachang), and used microscopic method for morphological identification, molecular method for amplifying a partial region of mcyB gene and ELISA method for microcystin analysis. In the present study, 80% of M. aeruginosa strains contained mcy gene, followed by 45% (10 strains) of M. icthyoblabe, 33% (1 strain) of M. wesenbergii, and 11% (4 strains) of M. flos-aquae. Each percentage of mcy gene in Microcystis morphospecies was similar to that of microcystin production in Microcystis morophospecies. In conclusion, the present study shows that molecular method using mcy gene primers can be used as an indirect indicator for the monitoring of toxic cyanobacteria (Microcystis).

Generation of Transgenic Plant (Nicotiana tabacum var. Petit Havana SR1) harboring Bacillus thuringiensis Insecticidal Crystal Protein Gene, cry II A (Bacillus thuringiensis 살충성 결정단백질 유전자(cry II A)의 형질전환 식물 제작)

  • 이정민;류종석;권무식
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.5
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    • pp.305-311
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    • 1997
  • Bacillus thuringiensis, a gram-positive soil bacterium, is characterized by its ability to produce crystalline inclusions during sporulation. The crystal proteins exhibit a highly specific insecticidal activity. An insecticidal crystal protein (ICP), Cry II A, is specifically toxic to both lepidopteran and dipteran insects. In this study, tobacco plants transformed by the cry II A gene have been generated. The Cry II A crystal protein was purified from E. coli JM103 harboring cry II A gene by differential solubility. The activated Cry II A was prepared by tryptic digestion. The purified protoxin (70 kDa) and the activated toxin (50 kDa) were analyzed by SDS-PAGE. To generate the transgenic tobacco having cry II A gene, the cry II A gene was subcloned to a plant expression vector, pSRL2, having two CaMV 35S promoters. The recombinant plasmid was transformed into tobacco (N. tabacum var. Petit Havana SR1) by Agrobacterium-mediated leaf disc transformation. Through the regeneration, six putative transgenic tobacco plants were obtained and three transformants were confirmed by Southern blot analysis. It has been found that one plant had single copy of cry II A gene, another had two copies of the gene, and the third had a truncated gene. After the immunochemical confirmation of cry II A expression in plants, the transgenic tobacco plants will be used to study the genetics of future generation with the insecticidal crystal protein gene cry II A.

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Toxins and Antibiotic Resistance of Methicillin-Resistant Staphylococcus aureus Isolated from Clinical Specimens (임상검체로부터 분리된 methicillin 내성 Staphylococcus aureus의 독소 및 항생제 내성)

  • Baik, Keun-Sik;Ki, Gwang-Seo;Choe, Han-Na;Park, Seong-Chan;Koh, Eun-Cho;Kim, Hyung-Rak;Seong, Chi-Nam
    • Journal of Life Science
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    • v.21 no.2
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    • pp.257-264
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    • 2011
  • Seventy five methicillin- resistant Staphylococcus aureus (MRSA) strains and 24 methicillin- susceptible S. aureus (MSSA) were isolated from clinical specimens obtained from a hospital in Suncheon, Jeonnam province, Korea, from July to December, 2009. Antibiotic resistance was determined using the disc diffusion method. Genes encoding enterotoxin (SE), toxic shock syndrome toxin-1 (TSST-1), exfoliative toxin (ET) and Panton-Valentine leukocidin (PVL) were detected by multiplex PCR-mediated amplification using specific primers. Sixty (80%) MRSA isolates possessed either one or more toxin genes and the most common pattern that coexisted in MRSA was seb, sec, seg, sei and tst (22.7%) followed by coexistence of sec, seg, sei and tst genes (18.7%). Gene pvl encoding leukocidin was not found. Significant correlation between the production of sec, seg, sei and tst genes was found. MRSAs were resistant to erythromycin (89% of the isolates), gentamicin (70.7%), ciprofloxacin (69.3%), clindamycin (61.3%) and tetracycline (58.7%), while MSSAs were susceptible to the antibiotics with the exception of erythromycin. Toxin genes seb, sec and tst were related to the tetracycline resistance of MRSA.

Evaluation of Nutritional Safety for the Herbicide-Resistant Rice in Growing Male Rats (성장기 흰쥐에서 제초제 저항성 쌀의 급여에 대한 영양적 안전성 평가)

  • 이성현;박홍주;조소영;전혜경;박용환;정미혜;박선희
    • Journal of Nutrition and Health
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    • v.36 no.10
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    • pp.1030-1035
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    • 2003
  • This study was conducted to evaluate the safety of herbicide-resistant rice, a genetically modified organism (GMO) developed by the Rural Development Administration, in Sprague-Dawley rats. The nutrient content of herbicide-resistant polished and brown cooked rice was compared with that of conventional Ilpum polished and brown cooked rice to assess composition equivalence. Compositional analysis was performed to measure proximates, fiber, and minerals before animal feeding. Growing male rats were fed one of the following four diets for six weeks: Ilpum polished cooked rice (IP) and Ilpum brown cooked rice (IB) as a non-GMO and herbicide-resistant polished cooked rice (GP) and brown cooked rice (GB) as a GMO. We checked clinical symptoms (anorexia, salivation, diarrhea, polyuria, anuria, fecal change) every day, food intake, change of body weight twice a week, and serum biochemistry and organ weights after 6 weeks of experimental feeding among the four groups. Nutrient content of the herbicide-resistant rice was similar to that of the non-transgenic control and was within the published range observed for non-transgenic rice. We could not find any significant difference in the above-mentioned items as the index to be checked in the animals fed the GMO. These results suggest that the nutrient content of genetically modified herbicide-resistant rice is compositionally equivalent to that of conventional Ilpum rice and that growing male rats fed herbicide-resistant rice are no different from those fed Ilpum rice, non-GMO for 6 weeks.

Trends in Protein Engineering for Gene Targeting: Homing Endonucleases and Zinc Finger Nucleases (유전자 표적화를 위한 단백질공학 연구동향: Homing Endonucleases and Zinc Finger Nucleases)

  • Cheong, Dea-Eun;Kim, Geun-Joong
    • KSBB Journal
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    • v.25 no.3
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    • pp.215-222
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    • 2010
  • Monogenic diseases are resulted from modifications in a single gene of human cells. Because their treatment with pharmacological medicine have a temporary effect, continuous nursing care and retreatment are required. Gene therapy, gene targeting and induced pluripotent stem cell (iPSC) are considered permanent treatment methods of them. In gene therapy, however, retroviral vectors that have potential toxicity caused by random insertion of harmful virus are used as vehicles for transferring genetic materials. On the other hand, gene targeting could replace and remove the modified gene though homologous recombination (HR) induced by site-specific endonucleases. This short review provides a brief overview on the recently tailored endonucleses with high selectivity for HR.