• Title/Summary/Keyword: 유전자독성

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Virulence Genes of Staphylococcus aureus Isolated in Daegu and Gyeongsangbuk-do Areas (대구광역시와 경상북도 지역에서 분리한 Staphylococcus aureus 병독소 유전자의 분자적 연구)

  • Jeon, Seok-Jae;Lee, Hee-Moo
    • Korean Journal of Clinical Laboratory Science
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    • v.40 no.1
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    • pp.48-54
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    • 2008
  • Nine types of staphylococcal enterotoxin (SE) genes (sea~see, seg~sej), 3 types of virulence genes (eta, etb, tst), mecA and 16S rRNA as internal positive control were detected from 187 clinical MRSA (methicillin resistance Staphylococcus aureus) strains isolated from a variety hospitalized patients in Daegu and Gyeongsangbuk-do areas using the multiplex PCR. The frequency of the S. aureus strains harboring recently reported SE genes (seg~sej) were found to be very high (65.9%) and greater than that of the strains harboring classical SE (sea~see) genes (47.8%) as previously established. Taking into account that the newly described pairs form SE genes (i.e., sec+seg+sei, seg+sei) were many, in the other hand, single form SE genes (i.e., seg, seh, sei and sej) were rarely detected. The S. aureus with pairs form enterotoxigenic genes become more potentially toxigenic strains. Furthermore, this work indicated a systematic association between the seg and sei genes and their high incidence among the S, aureus strains, which suggests that these two SE's could be an important phylogenetic link among the staphylococcal enterotoxins.

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Effect of Houttuynia cordata Extracts on Tyrosinase Gene Expression (어성초(Houttuynia cordata) 추출물이 Tyrosinase유전자 발현에 미치는 효과 -연구노트-)

  • Chin, Jong-Eon;Cho, Nam-Chul
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.8
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    • pp.1284-1288
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    • 2005
  • The methanol extract of Houttuynia cordata repressed the expression of tyrosinase gene of Bl6 mouse melanoma cell containing tyrosinase promoter. The extract repressed expression of tyrosinase gene by about 13$\%$ and 45$\%$ at 10$\mu$g/mL and 100 $\mu$g/mL, respectively. In the MTT assay, the same extract exhibited very low cytotoxicity at 1$\mu$g/mL, 10$\mu$g/mL, and 100 $\mu$g/mL, respectively. The fractions of ethyl acetate, butyl alcohol, and water did not showed the repressive effect on the expression of tyrosinase gone, but methylene chloride fraction layer repressed highly at 10$\mu$g/mL and 100$\mu$g/mL.

Characterization of gp64 Gene of Bombyx mori Nucleopolyhedrovirus and Development of a Transient Expression Vector (누에 핵다각체병 바이러스 헤 gp64 유전자의 특성조사 및 transient 발현 벡터 개발)

  • 김미향;최재영;우수동;이해광;제연호
    • Microbiology and Biotechnology Letters
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    • v.29 no.1
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    • pp.18-24
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    • 2001
  • Expression of the baculovirus major envelope glycoprotein gene(gp64) is regulated by transcription from botha early and late promoters. To develop a transient expression vector under the control of gp64 gene promoter, the gp64 gene of Bombyx mori nucleopolyhedrovirus-K1(BmNPV-K1) was characterized. The gp64 gene was local-ized at EcoR I-Pst I 7.38-kb fragment of the BmNPV-K1 genome. The EcorR 1-Pst I 7.38-kb fragment was cloned and the nucleotide sequence of 2,277 bases including the coding region of gp64 gene was determined. Based on these results, transient expression vector using gp64 gene promoter was constructed and named as pBm64. E.coli lacZ gene was introduced onto pBm64 as a reporter gene and expressed transiently in B. mori 5(Bm 5) cells. The expression vector transfected into the cells was maintained stably for 1 to 5 days. In order to confirm the expression of the reporter gene by gp64 promoter, recombinant virus was constructed. The recombinant virus has two independent transcription units in opposite orientations with two promoters; gp64 and polyhedrin gene promoters each initiating transcription of $\beta$-galactosidase and polyhedrin, respectively. Polyhedra formation and expression of $\beta$-galactosidase in Bm5 cells infected with the recombinant virus were observed with phase contrast microscope and in situ staining.

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Transcriptional Modulation of Metabolism-Related Genes in Brackish Water Flea Diaphanosoma celebensis Exposed to Mercury (수은 노출에 따른 기수산 물벼룩의 대사 관련 유전자의 발현 양상)

  • Min Jeong, Jeon;Je-Won, Yoo;Young-Mi, Lee
    • Journal of Marine Life Science
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    • v.7 no.2
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    • pp.145-153
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    • 2022
  • Mercury (Hg) is a major concern in marine environment because of their bioaccumulation and biomagnification properties, and adverse effects to aquatic organisms at even a trace amount. However, little information on the effects of Hg, compared to other heavy metals, is available in marine small crustaceans. Here, we investigated the transcriptional modulation of metabolism-related genes in the brackish water flea, Diaphanosoma celebensis after exposure to sublethal concentration (0.2, 0.4, 0.8 ㎍/l) of HgCl2 for 48 h. Relative mRNA expression levels of five detoxification enzyme-coding genes (cytochrome P450; cyp360A1, cyp361A1, cyp4AP3, cyp4C122, and cyp370C5) and six digestive enzyme-coding genes [alpha amylase (AMY), alpha amylase related protein (AMY-like), trypsin (TRYP), chymotrypsin-like protein (CHY), lipase (LIP), pancreatic lipase-related protein (PLRP)] were analyzed using quantitative real time reverse transcription polymerase chain reaction (qRT-PCR). As results, Hg increased the mRNA level of cyp370C5 (clan2) and cyp4AP3 (clan4) in a concentration dependent manner. A significant increase in TRYP mRNA was also concentration-dependently observed after exposure to Hg. These findings suggest that cyp370C5 and cyp4AP3 play a key role in Hg detoxification in D. celebensis, and Hg can affect energy metabolism by modulating the transcription of digestive enzyme. This study will provide better understanding the molecular effects of Hg in marine small crustacean.

파푸아뉴기니아산 열대 식물의 항변이원성 연구

  • 장일무
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.126-126
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    • 1993
  • 암을 유발시키는 원인은 생물, 물리, 화학적 요인 등 여러 가지가 있으나 근자에 들어 산업후에 따르는 환경오염의 증가는 화학적 원인에 의한 발암가능성을 증대시키고 있다. 이러한 화학적 인자의 발암 기전 중 많은 부분이 유전자에 상해를 일으킴으로써 암을 발생시킨다고 알려져 있다. 그러므로 항암실험을 수행하였던 파푸아뉴기니아산 열대식물 13과 23종 부위별 36시료에 대하여 항변이원성 검색실험을 통하여 유전독성을 가진 화학적 요인에 의한 발암 가능성에 대한 예방 효과를 알아봄으로써 발암에 대한 예방약물 개발의 기초 자료를 제공하고자 본 실험을 수행하였다.

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Tributyltin chloride(TBTC)와 Triphenyltin chloride(TPTC)에 노출된 넙치(Paralichthys olivaceus) 치어에 있어서의 RT-PCR에 의한 유전자 증폭반응

  • 최윤실;함승협;진덕희;전중균
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.373-374
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    • 2000
  • Tributylin(TBT)과 Triphenyltin(TPT) 같은 유기주석화합물은 선박, 어망, 어구등 방오도료의 소재로 사용되어 왔으나 최근 해양생물에게 미치는 독성 때문에 세계 각 국에서도 그 사용을 금지하고 있다. 우리 나라의 경우, 한국 해양연구소가 내분비 교란물질(환경호르몬)의 농도를 조사한 결과 다른 지역에 비해 내만 지역에서의 오염도가 다소 높다고 보고되어 있다. 내만지역의 경우는 파도가 약하여 양식장이 많으므로 양식어류가 TBT와 같은 유기주석 화합물에 노출될 가능성은 매우 높다. (중략)

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Inheritance Study of Male Sterile Transformants Containing Pollen-specific Promoter and Diphtheria Toxin A Gene (수술특이프로모터와 디프테리아 독성 유전자에 의한 웅성불임 형질전환체의 후대 유전분석)

  • Park, Young-Doo;Kim, Hyun-Uk;Park, Beom-Seok;Jin, Yong-Moon
    • Horticultural Science & Technology
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    • v.18 no.3
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    • pp.342-347
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    • 2000
  • The objectives of this study were to investigate the genetic and phenotypic features of male sterile transformants by pollen-specific expression of diphtheria toxin gene and to find out inheritance patterns of transgene to the next generation. When backcrossed (BC) progenies were tested for expression of kanamycin resistance ($Km^R$), 9 lines out of 13 lines, except 4 lines ($BC_{1}5-13,\;BC_{1}5-23,\;BC_{1}5-28,\;BC_{1}5-32$), showed the ratio of $Km^R$ to kanamycin sensitive ($Km^S$), from 1:30 to all $Km^S$. As a result, they were much lower than Mendelian segregation of a dominant gene. To determine whether male sterility is a heritable and stable trait, 5 male sterile plants ($BC_{1}5-13,\;BC_{1}5-14,\;BC_{1}5-23,\;BC_{1}5-32,\;BC_{1}5-33$ lines) which had different transgene copy numbers were backcrossed as female parents with pollens from wild type. To confirm the existence of the DTx-A gene in the genome of the progenies, PCR was conducted using specific primers of the DTx-A coding region. A PCR band of 428 bp was obtained from each generation, which is the predicted size of the DTx-A gene fragment. Trangenes were inherited to the next $BC_4T_0$ progenies and showed male sterility, however, based on the copy numbers of DTx-A gene male sterile plants did not show predicted ratio. When male sterile plants were backcrossed with fertile plants, fruit capsule sizes and seed settings were relatively reduced from those of selfing wild type plants. The fruit sizes and seed settings were reduced in proportion to the increase in the copy number of DTx-A gene.

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The Effects of Pro-inflammatory Cytokines by Cisplatin on the Death of Sensory Hair Cells. (시스플라틴에 의한 염증성 사이토카인의 청각유모세포 사멸 효과)

  • Lee, Jeong-Han;Park, Chan-Ny;Park, Rae-Kil
    • Journal of Life Science
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    • v.18 no.4
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    • pp.542-549
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    • 2008
  • Cisplatin (cis-diamminedichloroplatinum II : CDDP) is the most widely used anticancer drug against a variety of human neoplasms. However, its clinical use is limited by the onset of severe side effects, including ototoxicity and nephrotoxicity. Even though a number of evidences in cytotoxic mechanism of cisplatin have been suggested, the role of pro-inflammatory cytokines in cisplatin cytotoxicity of auditory cells has not yet been demonstrated. Herein our data clearly demonstrated that cisplatin decreased the viability of HEI-OC1 auditory cells, which was inhibited by the addition of neutralizing $anti-TNF-{\alpha}$, $anti-IL-1{\beta}$ and anti-IL-6 antibodies. Consistently, Neutralization with antibodies against pro-inflammatory cytokines ameliorated the cell death and disarrangement of cochlea hair cell layers in the rat primary cochlear explants which were treated with cisplatin. Furthermore, exogeneous supplementation with free radical scavengers, including GSH and NAC, significantly prevented the cytotoxicity of cisplatin in the rat primary cochlea explants. We also observed that $TNF-{\alpha}$ was predominantly expressed in Deiters and Hensen's cells located in hair cell zone of cisplatin-treated cochlear explants. These findings suggest that pro-inflammatory cytokines, including $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6, may play a pivotal role in the pathophysiology of hair cell damages caused by ototoxic drug cisplatin.

Conditions for Stable light Production of Recombinant Escherichia coli Containing Lux Operon and Sensitivity to Toxic Chemicals (Lux operon을 함유한 유전자 재조합 Escherichia coli의 발광 안정화 조건 및 독성물질에 대한 민감성)

  • 배희경;이상민;정윤철;송방호;신평균
    • KSBB Journal
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    • v.17 no.6
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    • pp.571-576
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    • 2002
  • Recombinant E. coli DH5 ${\alpha}$/pSB311 was made by cloning the genes encoding bacterial luciferase and aldehyde substrate proteins from Photohabdus luminescense, to complement defects of Lumistox, which is normally used in bioassays to monitor toxic substances in water environmental systems. The conditions for stable light production by the recombinant strains were investigated with respect to cell growth stage, cell number, and buffer conditions. The optimum growth stage was a middle-exponential stage with an OD$_{660nm}$ value of 0.6-0.7. ADout 10$^{6}$-10$^{7}$ cells per test tube was optimum for stable light emission. The effect of buffer was not significant if an optimum viable cell number was maintained. The bioluminescence of the recombinant E. coli harboring the lux operon of Photohabdus luminescense was not affected by temperature, while the bioluminescence of Lumistox was temperature sensitive. The recombinant E. coli was more sensitive to heavy metals (Cd, Cu, Hg, Zn) than Lumistox, because it does not require high concentrations of NaCl in the buffer.

Responses of Misgurnus anguillicaudatus and Cyprinus carpio Fed on Disease Resistant(OsCK1) Rice Variety (병저항성 GM(OsCK1)벼가 미꾸리(Misgurnus anguillicaudatus)및 잉어(Cyprinus carpio)에 미치는 영향)

  • Oh, Sung-Dug;Lee, Kijong;Park, Soo-Yun;Lee, Dae-Yong;Sohn, Soo-In;Kim, Min-Young;Ryu, Tae-Hun
    • Korean Journal of Environmental Agriculture
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    • v.32 no.3
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    • pp.231-239
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    • 2013
  • BACKGROUND: The disease resistant (OsCK1) rice was generated by inserting choline kinase (CK1) and phosphinothricin acetyltransferase (PAT) genes isolated from Oriza sativa and Streptomyces hygroscopicus into the genome of rice (Nakdongbyeo). With the potential problems of safety, the non-target organism evaluation is required as an essential element for the environmental risk assessment of genetically modified (GM) crops. In present study, we studied the effects on survival of Misgurnus anguillicaudatus and Cyprinus carpio, commonly used as a model organism in ecotoxicological studies. METHODS AND RESULTS: The M. anguillicaudatus and C. carpio were fed on disease resistant (OsCK1) rice and non-genetically modified (non-GM) rice (Nakdongbyeo) to 0, 10, 100, 1,000 and 5,000 mg/L, as treatment concentration respectively. The OsCK1 rice used for the test was confirmed to have the OsCK1/PAT gene expression by the PCR and ELISA analysis. Feeding test showed that no significant differences in cumulative immobility and abnormal response of M. anguillicaudatus and C. carpio fed on between OsCK1 rice and non-GM rice. The 96hr-$LC_{50}$ values showed no difference between OsCK1 rice (>5,000 mg/L) and non-GM rice (>5,000 mg/L). CONCLUSION(S): The results of this study suggested that there was no significant difference in toxicity for M. anguillicaudatus and C. carpio between OsCK1 rice and non-GM counterparts.