• International Journal of Industrial Entomology and Biomaterials
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• 제9권2호
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• pp.155-165
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• 2004

In Korean Peninsula including neighboring islands and Japanese Islands identical firefly species or the species belonging to same genera occur together in both territories. These geographic firefly species, nonetheless, have never been subject to taxonomic consideration together until recently, lacking clear species status and phylogenetic relationships. A recent serial study of these fireflies using luciferase gene and/or portions of mitochondrial DNA sequences provided some insight into these populations in terms of validity of species name, phylogenetic relationships, and speciation event. In this article, thus, we have reviewed the recent progress on phylogenetic and/or population genetic aspects of these species, i.e., Hotaria-group fireflies, Luciola lateralis, and Pyrocoelia rufa to better understand the firefly species in these regions.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 Molecular and Cellular Response to Toxic Substances
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• pp.164-164
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• 2002

The estrogenic activity of 47 plant extracts was assessed by reporter gene assay using MCF-7 breast cancer cell lines stably transfected with luciferase reporter gene. The estrogenic activity of food extracts was expressed as 17${\beta}$-estradiol(E2) equivalent concentration(EEQ), the concentration of E2 that resulted in the same relative luciferase unit(RLU) of the food extract of 0.2mg/$m\ell$.(omitted)

• Journal of Acupuncture Research
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• 제21권6호
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• pp.19-36
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• 2004

Objective : The purpose of this study was to investigate the effect of Bee Venom and Melittin Solution on the lipopolysaccharide(LPS) and sodium nitroprusside(SNP)-induced expression of prostaglandin $E_2(PGE_2)$, cyclooxygenase-2(COX-2), nuclear factor kappa B($NF-{\kappa}B$) and nuclear factor kappa B($NF-{\kappa}B$) dependent luciferase activity in RAW 264.7 cells, a murine macrophage cell line. Methods : The expression of PGE2 was determined by determination of $PEG_2$, COX-2 was by western blotting with corresponding antibodies, $NF-{\kappa}B$ was by gel mobility shift assay method and $NF-{\kappa}B$ dependent luciferase activity was investigated by luciferase assay in RAW 264.7 cells. Results : 1. LPS and SNP-induced expression of $PEG_2$ was significant after 24hour. 2. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS-induced expression of $PEG_2$ and, the $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly SNP-induced expression of $PEG_2$ compared with control, respectively. The 0.5 and $1{\mu}g/mL$ of bee venom could not significantly inhibit SNP-induced expression of $PEG_2$ compared with control. 3. The $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS and SNP-induced expression of COX-2 compared with control, respectively. The 0.5 and $1{\mu}g/mL$ of bee venom inclined to decrease LPS and SNP-induced expression of COX-2 compared with control. 4. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS and SNP-induced expression of $NF-{\kappa}B$ compared with control, respectively. 5. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS-induced expression of $NF-{\kappa}B$ dependent luciferase activity and the 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly SNP-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control, respectively. The $NF-{\kappa}B$ inhibitor also inhibited significantly LPS and SNP-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control. 6. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS + IFN-${\gamma}$, TNF-${\alpha}$ and LPS + TNF-${\alpha}$-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control, respectively. The $NF-{\kappa}B$ inhibitor also inhibited significantly LPS and SNP-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control. Conclusions : These results suggest the inhibitory action of bee venom and melittin solution on the inflammatory mediators such as $PEG_2$, COX-2 and $NF-{\kappa}B$.

• 농약과학회지
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• 제8권2호
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• pp.95-102
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• 2004

현재 국내 사용중이며 내분비계 장애추정농약으로 분류된 benomyl, carbaryl, endosulfan등 17종 농약에 대한 estrogen성 영향을 검색하기 위하여 인체난소암세포(BG1Luc4E2)를 이용한 luciferase assay를 수행하였으며, luciferase assay에서 Eeq를 산출한 후 내분비계 장애추정 농약의 에스트로겐성 영향에 대한 식이섭취 위험도 평가를 실시하였다. Estrogen 수용체 결합시험에서 cypermethrin, dicofol, endosulfan, esfenvalerate 및 fenvalerate가 $10^{-5}$ M에서 최고 영향이 관찰되었고, mancozeb 등 8종 농약은 약한 영향이 관찰되었으며, benomyl 등 나머지 4종 농약은 영향이 없었다. 이들 중 활성이 비교적 강한 dicofol 및 endoeulfan의 1 nmol 17 $\beta$-estradiol에 대한 RLP 와 RLU는 dicofol의 경우 $10^{-5}$ 및 56%이었구, endosulfan은 $10^{-5}$ 및 72%이었다. MRL을 이용한 식이섭취 위험도 평가 결과 농약들의 추청 1일 최대농약섭취량은 cypermethrin 0.667, dicofol 0.1462, endosulfan 0.2066 및 lenvalerate/esfenvalerate 0.2098 mg/person으로 총 추정 1일 최대 농약섭취량이 1.2298 mg/person이었고, 남성 혈중 에스트로겐 증가 농도는 3.075 ng/L로 정상농도에 비해 15%정도 증가하였으나, 국내 모니터링 성적을 기준으로 평가한 결과 남성혈중 에스트로겐 증가 농도는 0.01938 ng/L로 정상농도에 비해 0.09693%정도 증가하였다.

• Clinical and Experimental Pediatrics
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• 제48권5호
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• pp.495-499
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• 2005

목 적 : IL-4는 Th2 면역 반응의 중요한 매개체로 IL4 유전자의 promoter 일배체형(haplotype)은 한국인 소아에서 RSV에 의한 심한 모세기관지염과 연관된다고 알려져 있다. 본 연구는 IL4 유전자의 promoter 다형성에 따른 IL-4 단백의 기능적인 변화를 분석하여 심한 RSV 하기도 감염증에 기여하는 IL4 유전자의 발병 기전의 연관성을 연구하고자 시행하였다. 방 법 : 면역 기능이 정상인 소아 20명을 대상으로 전혈을 채취한 후 genomic DNA를 추출하여 IL4 유전자 promoter 약 1.2 kb 부위를 증폭시켰다. 염기서열 분석을 통하여 IL4 유전자 promoter의 유전형을 결정하고, PHASE 분석으로 일배체형을 결정하였다. 각 일배체형별로 $5{\mu}g$의 DNA를 Jurkat T 세포에 핵형질변환 시켜서 정상 Jurkat T 세포와 PMA(50 ng/mL)로 자극한 세포에서의 luciferase 활성도를 분석하여 IL4 유전자 promoter의 전사능을 결정하였다. 결 과 : 한국인 소아의 일배체형은 3가지 유형 GCC(7%), TCC(17%), 및 TTT(76%)로 분포하였다. Jurkat T 세포의 절대 luciferase 활성도는 GCC형이 가장 낮았고 TTT형이 가장 높았다. GCC 일배체형을 기준으로 하여 나타낸 Jurkat T 세포의 상대 luciferase 활성도는 TCC형이 4.2배, TTT형이 5.3배로 증가되었다. PMA로 자극한 후에 측정한 각 일배체형의 luciferase 활성도 역시 GCC형에 비하여 TCC형이 3.0배, TTT형이 4.1배로 증가되어 자극하지 않은 세포에서와 유사한 활성도의 차이를 보였다. 결 론 : 소아의 심한 RSV 하기도 감염증과 연관된 IL4 유전자의 promoter 일배체형 TTT는 IL4 유전자의 promoter의 전사능을 증가시킴으로써 영아 및 소아의 RSV 하기도 감염증의 병인에 중요한 역할을 할 것으로 생각된다.

• 한국동물학회지
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• 제39권4호
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• pp.378-386
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• 1996

우리는 두 가지의 새로운 luciferase reporter plasmid를 개발하였는데 그 하나는 이 plasmid에 promoter조각을 삽입한 다음에 그 promoter의 활성을 측정하는 용도로 사용 될수 있고, 다른 하나는 매우 낮은 basal promoter 활성을 갖고 있기 때문에 진핵생물의 전사 조절인자의 연구에 도움이 될 수 있다. 후자의 reporter plasmid에는 17염기쌍의 initator 와 Spl,GAL4그리고 일부 Drosophila homeodomain protein의 결합우뷔에 해당하는 cis elements등을 들어 있다.그리고 tranciption termination을 촉진할 수 있는 signal을 initiator앞서 삽입하여 이런 reporter plasmid에 존재할 수 있는 cryptic promoter에서 시작된 transcript가 luciferase reporter cDNA로 진행되는 것을 방지하는 개선된 reporter plasmid를 제조하여 promoter활성을 Drosophila Schneider line 2 cells을 이용한 transient transfection assay방법으로 측정하였다. 여기에 사용한 termination 촉진 signal은 SV40 polyadenylation signal의 3연속 조각(AAA)과 tenscription termination signal이 포함된 것으로 믿어지는 mouse c-mos 유전자의 일부조각 (UMS)이다. 기대한으로 이 두가지의 signal을 삽입하였을 때 basal promoter활성이 최대한으로 감소하였으며 이 두 가지 signal을 삽입된 reporter plasmid를 사용하여 promoter의 활성을 보다 sensitive하게 측정하였다. 이 reporter plasmid는 Droiophlla melanogaiter뿐만 아니라 포유동물을 포함한 고등생물의 전사 조절인자 연구의 한 도구로 사용될 수 있을 것이다.

• 대한치주과학회:학술대회논문집
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• 대한치주과학회 2006년도 제46회 종합학술대회 연제초록
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• pp.115-115
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• 2006

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.588-591
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• 2000

Sphingoliped 대사의 율속효소인 serine palmitoyl transferase(SPT)와 acid ceramidase의 연구를 위하여 인간의 SPTII 유전자와 acid ceramidase 유전자의 5‘-upstream region을 얻었다. 사람의 대장암세포인 HT29 cell로부터 genomic DNA를 얻고 GenomeWalker kit를 이용하였으며 2690bp의 SPTII promoter와 2028bp 및 1034bp의 acid ceramidase promoter의 fragment들을 얻을 수 있었다. 이들 DNA 조각들을 T7Blue vector에 subcloning하여 sequencing하였으며 이들이 사람의 SPTII 및 acid ceramidase gene의 5’-upstream region임을 확인하였다. 동물세포에서의 promoter activity를 측정하기위하여 firefly luciferase를 reporter gene으로 하는 pGL2-enhancer vector와 pGL2-basic vector에 subcloning하였으며 pRL-TK vctor와 함께 HT29 cell 및 HepG2 cell에 cotransfection 시킨 후 luciferaseg활성을 측정한 결과 같은 양의 DNA로는 사람의 SPTII promoter와 acid ceramidase promoter는 pRL-TK에 비하여 transfection efficiency가 아주 낮았으며 promoter 연구를 위하여는 pRL-TK vector의 양을 1/100으로 줄이는 것이 적당하였으며 HT29 cell보다는 HepG2 cell에 더 높은 발현율을 보였다.

• 대한의생명과학회지
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• 제9권4호
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• pp.195-201
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• 2003

Aromatic hydrocarbons are of major concern among genotoxic chemicals due to their toxicity and persistence. Some microorganisms can utilize aromatic hydrocarbons as carbon and energy sources by inducing expression of catabolic operon(s). The XylR regulatory protein activates transcription of the catabolic enzymes to degrade BTEX (benzene, toluene, ethylbenzene, and xylene) from its cognate promoters, Pu and Ps upon exposure of the cells to the aromatic hydrocarbons. The activity of XylR on the promoters was previously monitored using luciferase luc reporter system. The xylR, its promoter Pr and the promoter Po for the phenolic compound catabolic operon were introduced upstream of firefly luciferase luc in the pGL3b vector to generate about 7.1 kb of pXRBTEX. Here E. coli harboring the plasmid was freeze-dried under various conditions to fin,d optimal conditions for storage and transport. The cell viability and luciferase activity were maintained better, when the cells were freeze-dried at -7$0^{\circ}C$ in the addition of the 10% skim milk or 12% sucrose. However, coaddition of protectants such as 10% skim milk plus 10% glucose or 12% sucrose plus 10% glucose, resulted in much better viability and bioluminescence activity compared with the effect of single addition of each protectant. In addition, it was shown that the freeze-dried cells maintained almost intact bioluminescent activities and cell viability for at least 1 week after freeze-drying. This work demonstrated that the properly freeze-dried recombinant bacterial cells could be utilized as a whole-cell biosensor for simple and rapid monitoring of BTEX in the environment.

• 한국환경성돌연변이발암원학회지
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• 제23권1호
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• pp.30-34
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• 2003

Recent industrial society has human widely exposed to PAHs (polynuclear aromatic hydrocarbons) that are comming from the incomplete combustion of organic material as wider spread environmental contaminants. Biological activities of PAHs are not known although PAHs are considered as carcinogens. Our laboratory have been studied the effect of PAHs in the mouse liver hepa 1 cells. In this study, we examined the mouse liver hepa-l cells as a new bioassay system to evaluate bioactivity of PAHs. We have selected 13 PAHs to examine bioassay using cyp1a1-luciferase reporter gene expression system where cyp1a1 1.6 Kb 5flanking region DNA was cloned in front of luciferase reporter gene and this plasmid was transfected into hepa 1 cells transiently. This cells then used for the study to observe the effect of PAHs. We demonstrated that PAHs induced the CYP1A1 promoter and 7-ethoxyresolufin O-deethylase (EROD) activities in a concentration-dependant manner. Some of PAHs showed stronger stimulatory effect on CYP1 gene expression than TCDD. Acenaphthene, anthracene, fluorine, naphthalene, pyrene, phenanthrene, carbazole were weak responders to cyp1a1 promoter activity stimulation and EROD induction in hepa 1 cells and these chemicals seemed to respond less to EROD than cyp1a1 promoter activity. Benz(a)anthracene, benzo(b)fluoranthene, benzo(k)fluoranthene, chrysene, and dibenzo(a,h)anthracene showed strong response to cyp1a1 promoter activity stimulation and also EROD induction in hepa 1cells. Results of dose response study suggested that four strong responding PAHs, such as benzo(a)anthracene benzo(k)fluoranthene, chrysene, and dibenzo(a, h)anthracene might be mediated through arylhydrocarbon receptor system in hepa1 cells.