• 대한치과교정학회지
• /
• 제21권1호
• /
• pp.97-111
• /
• 1991

It is the aim of this study to investigate the effects of sodium fluoride and sodium orthovanadate upon the proliferation and activity of the osteoblast (MC3T3-E1 cells). MC3T3-E1 cells were cultured in $\alpha-MEM$ containing $10\%$ FBS and various concentration of sodium fluoride and sodium orthovanadate was appended to serum free media. DNA synthesis was examined through the $[^3H]$ thymidine incorporation into DNA. Collagen synthesis was examined through the $[^3H]$ proline incorporation into collagenase digestible protein and noncollagen protein. The following results were drawn; 1. Sodium fluoride stimulated the DNA synthesis of osteoblast significantly in dose-dependent manner within the concentration from $2{\mu}M$ to $10{\mu}M$ (P < 0.005). 2. Sodium orthovanadate stimulated the DNA synthesis of osteoblast significantly in dose-dependent manner within the concentration from $2{\mu}M\;to\;8{\mu}M$, however showed diminution at $10{\mu}M$ (P < 0.001). 3. Sodium fluoride and sodium orthovanadate stimulated the percent collagen synthesis of osteoblast significantly in dose-dependent manner within the concentration from $5{\mu}M$ to $10{\mu}M$ (P < 0.001). 4. Sodium fluoride and sodium orthovanadate stimulated the noncollagen synthesis of osteoblast significantly in dose-dependent manner within the concentration from $5{\mu}M\;to\;10{\mu}M$ (P < 0.001). In conclusion, sodium fluoride and sodium orthovanadate stimulate the proliferation and activity of osteoblast by stimulation of DNA synthesis and collagen and noncollagen synthesis in osteoblast.

• 한국미생물·생명공학회지
• /
• 제51권1호
• /
• pp.99-108
• /
• 2023

대기 입자상물질(particulate matter, PM) 시료의 곰팡이 메타게놈 분석을 위해 DNA 추출 및 유전자 증폭 시 여러 문제가 발생한다. 본 연구에서는 PM 시료로부터 DNA를 추출하는 방법과 polymerase chain reaction (PCR)을 위한 프라이머 및 온도 조건의 최적화를 위하여 다양한 조건으로 실험하였다. 여러 조건에서 DNA 추출 여부를 비교 평가한 결과, bufffer와 proteinase K를 이용하여 20분 동안 화학적 세포 용해 처리와 bead beating 처리를 한 후 상용 DNA 추출 kit를 사용하면 DNA를 효율적으로 추출할 수 있었다. PCR 조건을 최적화하기 위해 ITS2 유전자 영역을 증폭할 수 있는 10개 조합의 프라이머를 이용하여 PCR을 수행한 결과, ITS3tagmix3/ITS4 조합의 프라이머로 annealing 온도 58℃로 하였을 때 증폭된 PCR 산물의 농도가 상대적으로 높았다. 이 조건에서도 PCR 산물의 농도가 낮은 경우에는 1차 PCR 산물을 주형 DNA로 사용하여 nested PCR을 수행하면 만족스러운 농도로 ITS2 유전자를 증폭할 수 있었다. 본 연구에서 도출한 조건으로 서울 대기 PM2.5를 포집한 필터 시료 15종을 대상으로 DNA 추출과 PCR을 수행한 결과 성공적으로 ITS2 유전자 증폭이 가능하였다. 본 연구에서 최적화한 방법은 대기 PM 시료의 곰팡이 메타게놈을 분석하고 해석하는 연구에 활용 가능하다.

• 대한예방한의학회지
• /
• 제12권2호
• /
• pp.51-59
• /
• 2008

Objectives : Buplueri Radix (BR), used medical plant in Korea traditional medicine, contains various compounds, including a series of triterpene saponins known as saikosaponins. We performed this study for the protective effect of BR against oxidative damage induced by 1,2,4-benzentriol(BT) in human lymphocytes. Methods : In order to investigate the protective effect of BR against carcinogens, genotoxicity induced by benzene metabolite, BT were performed using cytokinesis-block micronucleus(CBMN) assay and comet assay. Results : The frequency of micronucleus at 25, 50 and $100{\mu}M$ concentration of BT were $8{\pm}2.36$, $23{\pm}2.31$, $35{\pm}4.17$ respectively. In addition of BR with concentration of 25 and $50{\mu}g/mL$, MN frequencies were significantly decreased. According to comet assay, BT induced DNA damage in a dose-dependent manner at concentration of 10 and 50 while BT with BR treatment decreased DNA breakage. No genotoxicity was observed by BR($25{\sim}50{\mu}g/mL$) treatment alone on DNA breakage. Since BT can induce DNA damage through the generation of reactive oxygen species(ROS), we examined the level of ROS in human lymphocytes treated with BT and/or BR using DCF-DA, ROS-sensitive probe. The generation of ROS in BT-treated cells was also observed, and BR addition inhibited the level of BT-induced DNA damage. Conclusions : From above results it is suggested that BR could protect the cell and DNA from pro-oxidant effect by ROS by BT

• 한국식품영양학회지
• /
• 제30권4호
• /
• pp.673-680
• /
• 2017

Fludarabine, a chain terminating anti-cancer drug, is a purine analogue that causes DNA strand breaks in normal cells. In this study, we determined if A. melanocarpa and Korean red ginseng extract mixture reduce cytotoxicity of fludarabine. Treatment of HaCaT cells with $10{\mu}M$ of fludarabine for 24 hours decreased cell viability and increased DNA strand breaks. Treatment of A. melanocarpa and Korean red ginseng extract mixture for 24 hours increased cell viability as compared with single extract treatment. The protective effect of these extracts on cell activity increased in a concentration-dependent manner. DNA strand breaks induced by fludarabine decreased as concentration of extract mixture increased. p-H2AX level, a marker of DNA strand breakage, decreased depending on the concentration of extract mixture. The effect of mixed extract of A. melanocarpa and Korean red ginseng on DNA damage is due to the anti-oxidative effect of A. melanocarpa and signal transmission through glucocorticoid receptor upon binding of saponin of Korean red ginseng.

• 생태와환경
• /
• 제54권3호
• /
• pp.170-189
• /
• 2021

환경유전자(eDNA)는 다양한 환경(수중, 토양, 대기)에 존재하는 생물체로부터 유래된 유전물질을 의미한다. eDNA는 높은 민감도, 짧은 조사시간 등 많은 장점들이 존재하며 이로 인해 생물 모니터링 및 유해생물과 멸종위기 생물을 탐색하는 분야에 다양하게 활용되고 있다. 이러한 eDNA를 채집하기 위해서는 대상생물 및 대상유전자뿐만 아니라 현장 여과방법 및 eDNA 보존방법과 같이 매우 다양한 항목들을 고려해야 한다. 특히 환경에서 eDNA를 채집하는 방법은 eDNA 농도와 직결되는 항목으로서 적절한 채집방법을 사용하여 eDNA를 채집할 때 정확한 분석결과를 얻을 수 있다. 또한 현장에서 채집한 eDNA를 보존하고 추출하는 과정에서도 정확한 방법을 사용하였을 때 현장에 분포하는 eDNA의 농도를 정확하게 파악할 수 있다. 특히 eDNA 연구를 시작하는 연구자들에게 eDNA 분야는 초기 진입 장벽이 매우 높은 기술로서 이를 위한 기초 자료가 매우 절실하다. 본 연구에서는 본 연구는 eDNA가 수생태계를 연구하기 위한 도구로서 보다 널리 이용되며, eDNA를 이용하기 시작하는 연구자들에게 도움을 주고자 수생태계에서 eDNA를 채집하고 및 운반하는 방법과 실험실에서 eDNA를 추출하는 방법을 소개하고, 보다 간편하고 효율적인 eDNA 채집 도구와 방법을 제시하였다.

• BMB Reports
• /
• 제32권5호
• /
• pp.492-496
• /
• 1999

We have investigated the biochemical properties of the herpes simplex virus type 1 (HSV-1) DNA polymerase without the UL42 protein (Pol), purified from insect cells infected with a recombinant baculovirus containing the UL30 gene. BSA and DTT have inhibitory effects on dAMP incorporation. Pol showed a greater turnover rate of steady-state single nucleotide incorporation at 12 mM $MgCl_2$ than at 2 mM $MgCl_2$. However, it showed a greater processivity of DNA synthesis at lower $MgCl_2$ concentration (1 mM, 2 mM) than at a higher $MgCl_2$ concentration (12.5 mM). These results are consistent with a slow DNA dissociation at lower $MgCl_2$ concentrations. Pol does not incorporate a correct nucleotide into the primer with an incorrect nucleotide at the end; instead, it preferentially excises the incorrect nucleotide at the 3' end of the primer. Pol has DNA polymerase activity at pHs 6.5 and 7.5 but little at pHs 5.5, 8.5, and 9.5. It has exonuclease activity at pHs 6.5, 7.5, and 8.5 but little at pHs 4.5, 5.5, and 9.5. The finding that Pol has exonuclease activity but not DNA polymerase at pH 8.5 suggests that DNA binds to Pol, but deoxynucleotide binding or incorporation does not occur at pH 8.5.

• 한국수산과학회지
• /
• 제20권2호
• /
• pp.136-145
• /
• 1987

말똥 성게를 인공 수정시킨 후 column chromatography법으로 DNA polymerase $\alpha$를 분리 정제하였다. Sephadex G-200과 SDS polyacryamise gel electophoresis에 의한 DNA Polymerase $\alpha$의 분자량은 약 $137,000\~138,000$이였다. 효소활성의 최적 pH는 7.4였고, 칼슘이온 20mM, 나트륨이온 25mM에서 활성이 높았고, 마그네슘 이온은 10 mM 일 때 활성이 높았다. DNA polymerase $\alpha$는 N-ethylmaleimide, aphidicolin, cytosin $\beta-D-arabinofuranoside$ 5'-triphosphate (ara CTP)와 phosphonoacetic acid체 의하여 활성이크게 저하되었다.

• Journal of Oral Medicine and Pain
• /
• 제20권2호
• /
• pp.497-513
• /
• 1995

Human genomic Deoxyribonucleic acid(DNA) was extracted from teeth by boiling, salting-out, phenol, boiling-phenol methods. The author compared DNA concentration and its purity, the accuracy of sex determination and the results of the D1S80 locus detection among above 4 methods. The following results were obtained : 1. DNA concentration was the highest in pulp with salting-out method and DNA purity was higher in pulp with salting-out and phenol methods than other 2 methods. 2. Sex determination was possible using of the pulp and the dentin of the teeth with four methods but, it was impossible in the enamel and some pulp with boiling method. 3. Amplification of D1S80 locus occurred from pulp and dentin with salting-out, phenol, and boiling-phenol methods. 4. There are no differences among the amplification of X-Y homologus amelogenin gene by application of 4 methods and salting-out, phenol methods efficiently makes available to amplification of D1S80 locus. From the investigation DNA extraction, sex determination, amplification of D1S80 locus was successfully accomplished with salting-out, phenol, boiling-phenol methods Therefore above 3 methods are available and applicable as forensic odontology for individual identification.

• Journal of Radiation Protection and Research
• /
• 제41권4호
• /
• pp.339-343
• /
• 2016

Background: Evaluation of deoxyribonucleic acid (DNA)-strand break is important to elucidate the biological effect of ionizing radiations. The conventional methods for DNA-strand break evaluation have been achieved by Agarose gel electrophoresis and others using an electrical property of DNAs. Such kinds of DNA-strand break evaluation systems can estimate DNA-strand break, according to a molecular weight of DNAs. However, the conventional method needs pretreatment of the sample and a relatively long period for analysis. They do not have enough sensitivity to detect the strand break products in the low-dose region. Materials and Methods: The sample is water, methanol and plasmid DNA solution. The plasmid DNA pUC118 was multiplied by using Escherichia coli JM109 competent cells. The resonance frequency and Q-value were measured by means of microwave dielectric absorption spectroscopy. When a sample is located at a center of the electric field, resonance curve of the frequency that existed as a standing wave is disturbed. As a result, the perturbation effect to perform a resonance with different frequency is adopted. Results and Discussion: The resonance frequency shifted to higher frequency with an increase in a concentration of methanol as the model of the biological material, and the Q-value decreased. The absorption peak in microwave power spectrum of the double-strand break plasmid DNA shifted from the non-damaged plasmid DNA. Moreover, the sharpness of absorption peak changed resulting in change in Q-value. We confirmed that a resonance frequency shifted to higher frequency with an increase in concentration of the plasmid DNA. Conclusion: We developed a new technique for an evaluation of DNA damage. In this paper, we report the evaluation method of DNA damage using microwave dielectric absorption spectroscopy.

• Journal of Microbiology
• /
• 제44권5호
• /
• pp.508-514
• /
• 2006

In this study, the double-stranded DNA-dependent activities of Deinococcus radiodurans RecA protein (Dr RecA) were characterized. The interactions of the Dr RecA protein with double-stranded DNA were determined, especially dsDNA-dependent ATP hydrolysis by the Dr RecA protein and the DNA strand exchange reaction, in which multiple branch points exist on a single RecA protein-DNA complex. A nucleotide cofactor (ATP or dATP ) was required for the Dr RecA protein binding to duplex DNA. In the presence of dATP, the nucleation step in the binding process occurred more rapidly than in the presence of ATP. Salts inhibited the binding of the Dr RecA protein to double-stranded DNA. Double-stranded DNA-dependent ATPase activities showed a different sensitivity to anion species. Glutamate had only a minimal effect on the double-stranded DNA-dependent ATPase activities, up to a concentration of 0.7 M. In the competition experiment for Dr RecA protein binding, the Dr RecA protein manifested a higher affinity to double-stranded DNA than was observed for single-stranded DNA.