• Title/Summary/Keyword: 유전자독성

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Expressed Sequence Tag Analysis of Toxic Alexandrium tamarense and Identification of Saxitoxin Biosynthetic Genes (독성 Alexandrium tamarense 의 EST 분석 및 삭시톡신 생합성 유전자의 확인)

  • Chang, Man;Lee, Juyun;Chung, Youngjae;Lee, Gunsup;Kim, Dongguin;Lee, Taek-Kyun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.7
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    • pp.3582-3588
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    • 2013
  • Expressed sequence tag (EST) library was constructed from A. tamarense. Base sequences of EST clones were analyzed and saxitoxin biosynthesis-related genes were cloned. Sequences of 827 clones were analyzed and 564 EST were functionally clustered using Blast searches against GenBank. Main genes in the EST had functions on cellular organization, cell metabolism, energy, cell cycle and DNA processing, cellular transport and transport, cell rescue, defense, death and aging, and transcription. Moreover, expression of S-adenosylmethionine synthetase and H2A histone family genes were increased in the toxic A. tamarense. These results show that two genes could be a good biomarkers for the detection of saxitoxin biosynthesis in the A. tamarense.

Characterization of Synechocystic PCC6803 transformed with cryIVD gene of Bacillus thuringiensis subsp. morrisoni PG-14 and its mosquitochidal effect on Anopheles sinensis (Bacillus thuringiensis subsp. morrisoni PG-14 cryIVD 유전자로 형질전환된 Synechocytis PCC6803의 특성과 학질모기에 대한 살충효과)

  • 이대원;박현우;김호산;진병래;유효석;김근영;강석권
    • Korean journal of applied entomology
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    • v.35 no.1
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    • pp.66-73
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    • 1996
  • For the effective control of mosquito larvae, Anopheles sinensis, the expression vector pCYASK5-1 containing cryIVD gene of Bacillus thuringiensis subsp. morrisoni PG-14 was constructed and transformed into the cyanobacterium Synechocystis PCC6803. The transformants were selected on BG-11 medium containing kanamycin. The expression of cryIVD gene in transformant was confirmed by SDS-PAGE and Western blot analysis. The mortality of A. sinensis larvae was scored for 3 days. Furthermore, growth and distribution rate of transformant were examined. The results showed that Synechocystis PCC6803 transformed with cryIVD gene of B. thuringiensis subsp. morrisoni PG-14 was highly toxic to A. sinensis larvae, demonstrating that it will be a potential agent for mosquito control.

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Analysis of Antimicrobial Resistance Pattern and Distribution of Multi-drug Efflux Pump Genes and Virulence Genes in Enterococcus faecalis Isolated from Retail Meat in Seoul (서울시내 시판 식육에서 분리한 Enterococcus faecalis의 항생제 내성 유형, 다중약물 유출 펌프 유전자 및 병독성 유전자의 분포도 분석)

  • Choi, MinKyung;Choi, SungSook
    • Journal of Food Hygiene and Safety
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    • v.32 no.2
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    • pp.135-140
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    • 2017
  • The aim of this study was to investigate the distribution of genes that encode multi-drug efflux pumps and virulence factors in Enterococcus faecalis isolated from retail meat and antibiotic resistance patterns of these strains. Of the 277 retail meat samples, 93 Enterococcus faecalis were isolated. The strains exhibited resistance to ampicillin (35.5%), chloramphenicol (6.4%), ciprofloxacin (4.3%), erythromycin (18.3%), levofloxacin (0%), quinupristin-dalfopristin (76.3%), tetracycline (45.2%), teicoplanin (0%) and vancomycin (0%). The strains were positive for MFS type eme(A) (100%), ABC type efr(A) (100%), ABC type efr(B) (98.9%) and ABC type lsa (91.4%) efflux pump gene. The strains were positive for gelE (68.8%), ace (90.3%), asa1 (47.3%), efaA (91.4%) and esp (12.9%) virulence gene. This research will help to assess the hazards associated with the occurrence of drug resistance among enterococci from retail meat. Therefore, it is necessary to monitor enterococcus spp. isolated from retail meat continuously.

Development of Exposure Biomarkers for Endocrine Disrupting Chemicals Using DNA Microarray (DNA 마이크로어레이를 이용한 내분비장애물질 노출지표 개발)

  • Yang, Mi-Hi
    • Environmental Analysis Health and Toxicology
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    • v.20 no.4 s.51
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    • pp.327-332
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    • 2005
  • 장기간 노출 시 발암 등 인체 유해성을 갖는 환경유래 내분비장애물질(endocrine disrupting chemicals, EDCs)에 대한 선택적이고 민감한 노출지표를 개발하기 위하여 본 연구에서는 DNA microarray를 이용하였다. 피험자는 아직 특별한 질환을 갖지 않는 18세 이상 연령, 성을 맞춘 EDCs고농도 노출군(N = 16)과 저농도군(N = 16)으로 구성되었다. 노출정도 구분은 10년 이상 거주지가 K산업폐기물 소각장과 2.5 km 반경 내, 외 인지에 따라 고노출군,저노출군으로 구분하였다. 피험자의 말초혈에서 total RNA를 분리, 각 군당 B인씩 pool로 cDNA를 합성하여 oligonucleotide DNA 칩에 적용하였다. 유전자발현의 차이를 GenePixPro 4.0 software를 이용하여 분석하였다. 총 3장의 칩을 이용하여 공통적으로 저노출군보다 고노출군에서 2배 이상 발현의 증가를 보인 유전자는 plasminogen activator(PLAT)등 12종이 관찰되었고, l/2이하로 발현의 감소를 보인 유전자는 kallikrein 3 (KLK3)등 29종이었다. 이 들 유전자는 PLAT등 면역계 반응에 관여하는 유전자 및 apoptosis, transport, G protein, chromatin, 암화, 발생 (development), 대사 등에 관여하는 유전자들이었다. 그러므로 KLK3등 본 연구에서 발굴한 유전자는 향후 확대된 인구에서 본 연구 결과의 확인을 통하여 EDCs특이적 노출지표로써, 나아가 암 등 EDCs관련 질병의 기전 및 병인학을 구명하는데 이용가치가 높다고 사료된다.

The Mechanisms of Resistance to TNF in TNF-Sensitive Cancer Cells Transfected with TNF-$\alpha$ Gene Using Retroviral Vector (암세포에서 Retroviral Vector를 이용한 종양괴사인자 유전자 이입후 획득된 종양괴사인자 내성의 기전)

  • Lee, Hyuk-Pyo;Oh, Yeon-Mok;Yoo, Chul-Gyu;Kim, Young-Whan;Shim, Young-Soo;Han, Sung-Koo
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.547-558
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    • 1997
  • Background : Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on its potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majorities of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to hoot. In previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-$\alpha$ cDNA would become highly resistant to TNF. Understanding the mechanisms of TNF-resistance in TNF-$\alpha$ gene transfected cancer cells would be an important step for improving the efficacy of cancer gene therapy as well as for better understandings of tumor biology. This study was designed to evaluate the role of new protective protein synthesis in the acquired resistance to TNF of TNF-$\alpha$ gene transfected cancer cells. Method : We transfected TNF-$\alpha$ c-DNA to WEHI164, a murine fibrosarcoma cell line, using retroviral vector(pLT12SN(TNF)) and confirm the expression of TNF with PCR, ELISA, MIT assay. Then we determined the TNF resistance of TNF gene transfected cells(WEHI164-TNF) and the changes of TNF sensitivities after treatments with actinomycin D(transcription inhibitor) and cycloheximide ( translation inhibitor). Results : WEHI164 which was sensitive to TNF became resistant to TNF after being transfected with TNF-$\alpha$ gene and the resistance to TNF was partially reversed after treatment with actinomycin D, but not with cycloheximide. Conclusion : The acquired resistance to TNF after TNF-$\alpha$ gene transfection may be associated with synthesis of some protective proteins.

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Potential Importance of Proteomics in Research of Reproductive Biology (생식생물학에세 프로테오믹스의 응용)

  • Kim Ho-Seung;Yoon Yong-Dal
    • Development and Reproduction
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    • v.8 no.1
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    • pp.1-9
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    • 2004
  • The potential importance of proteomic approaches has been clearly demonstrated in other fields of human medical research, including liver and heart disease and certain forms of cancer. However, reproductive researches have been applied to proteomics poorly. Proteomics can be defined as the systematic analysis of proteins for their identity, quantity, and function. It could increase the predictability of early drug development and identify non-invasive biomarkers of toxicity or efficacy. Proteome analysis is most commonly accomplished by the combination of two-dimensional gel electrophoresis(2DE) and MALDI-TOF(matrix-assisted laser desorption ionization-time of flight) MS(mass spectrometry) or protein chip array and SELDI-TOF(surface-enhanced laser desorption ionization-time of flight) MS. In addition understanding the possessing knowledge of the developing biomarkers used to assess reproductive biology will also be essential components relevant to the topic of reproduction. The continued integration of proteomic and genomic data will have a fundamental impact on our understanding of the normal functioning of cells and organisms and will give insights into complex cellular processes and disease and provides new opportunities for the development of diagnostics and therapeutics. The challenge to researchers in the field of reproduction is to harness this new technology as well as others that are available to a greater extent than at present as they have considerable potential to greatly improve our understanding of the molecular aspects of reproduction both in health and disease.

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Effect of Bisphenol A on Early Embryonic Development and the Expression of Glutathione S-transferase (GST) in the Sea Urchin (Hemicentrotus pulcherrimus) (말똥성게(Hemicentrotus pulcherrimus)의 초기배아 발생과 glutathione S-transferase (GST)의 발현에 대한 bisphenol A의 영향)

  • Hwang, Un-Ki;Kim, Dae-Han;Ryu, Hyang-Mi;Lee, Ju-Wook;Park, Seung-Yoon;Kang, Han Seung
    • Korean Journal of Environmental Biology
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    • v.32 no.3
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    • pp.234-242
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    • 2014
  • In this study, gametotoxicity and embryotoxicity experiments using Hemicentrotus pulcherrimus were carried out to investigate the ecotoxicological effects of bisphenol A (BPA). We examined the effects of BPA on fertilization and normal embryogenesis at various concentrations (0, 300, 500, 800, 1000, and 1500 ppb). The results demonstrated that the fertilization rates were not changed. The normal embryogenesis rates were gradually decreased in a dose-dependent manner, and were significantly lowered following 800 ppb BPA treatment ($EC_{50}$=1056.1 ppb, 95% Cl=981.8~1163.9 ppb). The observed effective concentration and the lowest observed effective concentration of the normal embryogenesis rate were 500 ppb and 800 ppb, respectively. The embryos showed retarded development at each tested concentration, indicating the fact the embryonic development was delayed due to the increasing concentrations of BPA. Furthermore, we examined the expression of glutathione S-transferase (GST) mRNA at various concentrations of BPA in H. pulcherrimus. Interestingly, it was found that the expression level of GST mRNA was significantly increased in the experimental group exposed to BPA. Based on these results, we suggested that BPA at greater than 800 ppb has a toxic effect during the early embryonic stages of H. pulcherrimus, and GST mRNA may be used as a biomarker for risk assessment of BPA contamination.

Functional Characterization of sll1556 of Synechocystis sp. PCC6803 as Type II Isopentenyl Diphosphate Isomerase (Type II Isopentenyl Diphosphate Isomerase로서 Synechocystis sp. PCC6803의 sll1556의 작용 특성)

  • Cho, Kab-Yeon
    • The Korean Journal of Food And Nutrition
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    • v.23 no.4
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    • pp.526-530
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    • 2010
  • Isopentenyl diphosphate(IPP) isomerization to dimethylallyl diphosphate(DMAPP) is an important step for the efficient production of isoprenoids such as lycopene, ${\beta}$-carotene, astaxanthin, etc. The type II isopentenyl diphosphate isomerase gene from Synechocystis sp. PCC6803(sll1556, Syidi2) was cloned and expressed in Escherichia coli $DH5{\alpha}$. When E. coli $DH5{\alpha}$ harboring lycopene synthesis genes, crtE, crtB, and crtI and mevalonate pathway genes, MvK1, MvK2, and Mvd, was cultured on LB medium containing mevalonate, the strain grew very slowly be due to the toxicity of isopentenyl diphosphate derived from mevalonate. When Syidi2 was introduced to E. coli $DH5{\alpha}$ harboring the lycopene synthesis genes and mevalonate pathway genes, growth on mevalonate medium was fully restored and the colony showed red color indicating lycopene formation. The growth rate of the mutant strain, E. coli $DH5{\alpha}$(idi::${\Delta}km$), was very slow because of IPP accumulation and DMAPP deprivation. Ultimately the idi mutant was complemented by introducing the Syidi2 gene.

Association Study Between the C3123A Polymorphism of the Angiotensin II Type 2 Receptor Gene in the Human X Chromosome and Essential Hypertension in Koreans (한국인에서 Angiotensin II Type 2 Receptor 유전자에 존재하는 C3123A 다형선과 본태성 고혈압과의 관련성에 관한 연구)

  • Kang Byung Yong;Bae Joon Seol;Lee Kang Oh
    • Environmental Analysis Health and Toxicology
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    • v.20 no.1
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    • pp.39-45
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    • 2005
  • Renin-angiotensin system (RAS)은 혈압 조절에 중요한 역할을 수행하는 생리적 조절계로써, 이 system 을 구성하는 유전자들의 이상은 본태성 고혈압의 발병과 유의하게 관련된 것으로 알려졌다. RAS의 주요한 구성 성분인 angiotensin II는 2종류의 수용체인 angiotensin II type I receptor(AT₁R)와 angiotensin II type I receptor(AT₂R)에 의해 그 효과가 매개되기 때문에, 이 수용체를 암호하는 유전자는 본태성 고혈압의 유력한 후보 유전자라고 볼 수 있다. 현재가지의 연구에 의하면, AT₁R 유전자에 존재하는 유전적 변이와 본태성 고혈압과의 관련성에 관해서는 많은 보고들이 있었지만, AT₂R 유전자에 존재하는 유전적 변이 가 본태성 고혈압에 유의한 효과를 나타내는 지에 관해서는 이렇다할 연구 성과가 별로 없는 실정이다. 이에 본 연구에서는 한국인 집단을 대상으로 하여, AT₂R 유전자에 존재하는 C3123A 다형성이 한국인 집단에서 본태성 고혈압과 유의한 관련성이 있는 지를 분석하였다. 이 유전자는 인간의 X 염색체에 존재하기 때문에, 여성인 경우에는 CC, CA및 AA로 이루어진 3유전자형이 존재하지만, 남성인 경우에는 C와 A로 이루어진 2종류의 대립 유전자로 구성되어 있기 때문에, 본 연구에서는 남성과 여성을 개별적으로 나누어서 분석하였다. 연구 결과, AT₂R 유전자에 존재하는 C3123A 다형성은 남녀 모두에서 본태성 고혈압과 유의한 관련성을 나타내지 않았다(P>0.05). 그렇지만, 이 다형성에 대한 대립 유전자 빈도를 서양인 집단과 비교했을 경우에는, 한국인을 대상으로 한 본 연구에서 A 대립 유전자 빈도가 0.33인 반면에 서양인 집단은 그 빈도가 0.43~0.48로 한국인 집단보다 높은 값을 나타내었다. 따라서, AT₂R 유전자에 존재하는 C3123A 다형성과 본태성 고혈압과의 관련성에 대해서는 한국인과 유전적 배경이 다른 서양인 집단을 대상으로 한 추시가 필요할 것으로 사료된다.

Bioassessment and Comparison of Toxicity of Arsenics based on the Results of Various Bioassays (다양한 생물 검정법에 근거한 비소의 위해성 평가 비교)

  • Kong, In-Chul;Kwon, Hyo-Jung;Ko, Kyung-Seok
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.8
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    • pp.795-801
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    • 2010
  • The acute toxicity of arsenic compounds was assessed and compared using following four bioassays; bioluminescence activity of the recombinant strain RB1436, germination of four different seeds, ${\alpha}$-glucosidase activity produced by Bacillus lichemiformis, acute genetic revertant mutation using mutant strain Salmonella typhimurium. Different sensitivities were observed among tested bioassays, but generally the toxicity by arsenite was greater than that of arsenate. Among tested four seeds, sensitivities of Lactucus and Raphanus were greater than others, and these two seed types were appeared as proper type for bioassay. High revertant mutation ratio (5.1) was observed with 1 mg/L arsenite, indicating high mutagenicity. The sensitivity of ${\alpha}$-glucosidase activity on arsenic compounds was much lower than other methods. The evaluation of interactive toxic effects using various bioassays may comprise a useful tool for the bioassessment of environmental pollutants.