• Title/Summary/Keyword: DNA 추출

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Protective Effect of Glycyrrhiza glabra Extract on UV-induced Skin DNA Damage (감초추출물(Glycyrrhiza glabra Extract)의 피부에서의 DNA 손상 방지효과)

  • Shin, Jae Young;Kang, Nae Gyu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.1
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    • pp.33-38
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    • 2022
  • Ultraviolet B (UVB) damages DNA residues in skin keratinocytes. In particular, the formation of cyclobutane pyrimidine dimers (CPD), a pyrimidine residue damage in DNA, is considered a representative indicator of skin photoaging. In this study, we confirmed defensive effect of Glycyrrhiza glabra (G. glabra) extract against UVB induced DNA damage. First of all, we confirmed UVB dependent amount of CPD formation in human keratinocyte cell line. UVB induced CPD was decreased by G. glabra extract by dose dependent manner. In addition, it was confirmed that the expression of mRNA of DNA damage recovery factors was increased by G. glabra extract. Consequently, through this study, it was possible to confirm the DNA protection effect of G. glabra extract in skin keratinocytes.

Optimization of DNA Extraction and PCR Conditions for Fungal Metagenome Analysis of Atmospheric Particulate Matter (대기 입자상물질 시료의 곰팡이 메타게놈 분석을 위한 DNA 추출 및 PCR 조건 최적화)

  • Sookyung Kang;Kyung-Suk Cho
    • Microbiology and Biotechnology Letters
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    • v.51 no.1
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    • pp.99-108
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    • 2023
  • Several challenges arise in DNA extraction and gene amplification for airborne fungal metagenome analysis from a particulate matter (PM) samples. In this study, various conditions were tested to optimize the DNA extraction method from PM samples and polymerase chain reaction (PCR) conditions with primer set and annealing temperature. As a result of comparative evaluation of DNA extraction under various conditions, chemical cell lysis using buffer and proteinase K for 20 minutes and bead beating treatment were followed by using a commercial DNA extraction kit to efficiently extract DNA from the PM filter samples. To optimize the PCR conditions, PCR was performed using 10 primer sets for amplifying the ITS2 gene region. The concentration of the PCR amplicon was relatively high when the annealing temperature was 58℃ with the ITS3tagmix3/ITS4 primer set. Even under these conditions, when the concentration of the PCR product was low, nested PCR was performed using the primary PCR amplicon as the template DNA to amplify the ITS2 gene at a satisfactory concentration. Using the methods optimized in this study, DNA extraction and PCR were performed on 15 filter samples that collected PM2.5 in Seoul, and the ITS2 gene was successfully amplified in all samples. The optimized methods can be used for research on analyzing and interpreting the fungal metagenome of atmospheric PM samples.

A Rapid Small Scale Method for Extraction of Genomic DNA from Lactobacillus spp. (Lactobacillus spp 로부터 Genomic DNA추출을 위한 신속/간단한 방법)

  • 이석용
    • KSBB Journal
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    • v.15 no.4
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    • pp.411-413
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    • 2000
  • A method is described for the rapid and simple isolation of genomic DNA from 3 mL culture of Lactobacillus crispatus KLB46 The isolated DNA using this method was shown to be an excellent substrate for restriction endonclease digestion and PCR. The method is expected to be used in gentic manipulation of L. crispatus KLB46.

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Effect of Salviae Radix on Cell Death and DNA Damage in Renal Proximal Tubular Cells Exposed to $H_2O_2$ ($H_2O_2$에 노출된 신장 근위 세뇨관 세포에서의 세포 사망 및 DNA 손상에 대한 단삼의 효과)

  • Sung-Dae, Kim;Ji-Cheon, Jeong
    • The Journal of Korean Medicine
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    • v.22 no.3
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    • pp.21-30
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    • 2001
  • 목적 : 이전 연구에서 단삼 추출액이 강력한 항산화 작용이 있음을 확인한 바 있어 단삼 추출액이 신장세뇨관 세포에서 oxidant에 의한 세포사망 및 DNA 손상을 방지하는 지를 조사하고 이러한 효과가 지질의 과산화를 억제하는 효과에 기인하는 지를 시험하였다. 방법 : 신장 근위세뇨관 세포 유래 세포주인 opossum kidney (OK)세포를 이용하여 세포 사망은 frypan blue exclusion방법을 이용하여 평가하였고, DNA손상 정도는 double stranded DNA의 파괴를 측정하여 평가하였다. Oxidant 약물 모델로는 $H_2O_2$를 사용하였다. 결과 : $H_2O_2$는 적용시 간과 농도에 비례하여 세포 사망을 유도하였다. 단삼 추출액은 0.05% 농도에서 $H_2O_2$에 의한 세포사망 및 DNA 손상을 방지하였다. 이러한 방지효과는 $H_2O_2$ 제거 효소인 catalase와 철 착염제인 deferoxamine에 의해서도 나타났다. 그러나 강력한 항산화제인 DPPD는 $H_2O_2$에 의한 세포 사망이나 DNA손상을 방지하지 못하였다. $H_2O_2$는 세포내 ATP 농도를 감소시켰으며. 이러한 감소는 poly (ADP-ribose) polymerase억제제인 3-aminobenzamide에 의해 방지되었으나 단삼 추출액에 의해서는 영향을 받지 않았다. 3-aminobenzamide는 $H_2O_2$에 의한 세포 사망을 방지하였다. $H_2O_2$는 지질의 과산화를 증가시켰으며, 이러한 변화는 단삼 추출액과 DPPD에 의해 방지되었다 결론 : OK 세포에서 $H_2O_2$에 의한 세포사망과 DNA 손상에는 지질의 과산화가 중요한 역할을 하지 않으며, 단삼 추출액의 $H_2O_2$에 의한 세포 사망과 DNA 손상 방지 효과는 항산화 작용이 아닌 다른 기전에 기인하는 것으로 사료된다.

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Inhibitory Effect of Methanol Extract of Doenjang on Growth and DNA Synthesis of Human Cancer Cells (된장 메탄올 추출물의 인체 암세포 성장 억제 효과 및 DNA 합성 저해 효과)

  • 임선영;이숙희;박건영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.6
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    • pp.936-940
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    • 2004
  • Doenjang (Korean soy paste) is one of the popular soybean based fermented foods in Korea. This study investigated the growth and DNA synthesis inhibitory effect of doenjang methanol extracts on AGS human gastric adenocarcinoma cells, Hep 3B human hepatocellular carcinoma cells and HT-29 human colon cancer cells. In order to determine an anticancer effect of doenjang methanol extracts, other soybean fermented foods and original materials were compared. The treatment of doenjang methanol extracts (200 $\mu\textrm{g}$/mL) to the AGS, Hep 3B and HT-29 cancer cells inhibited the growth of cancer cells by 80%, 77% and 86%, respectively. Compared to other soybean fermented foods and original materials, doenjang methanol extracts showed the highest growth inhibitory effect on different cancer cells. In addition, doenjang methanol extracts (200 $\mu\textrm{g}$/mL) significantly inhibited DNA synthesis of AGS and Hep 3B cancer cells by 76% and 59%, respectively. These results suggested that this anticancer effect of doenjang may be due to specific active compounds, which will be newly produced during soybean fermented process and not contained in soybean.

Modified CTAB DNA Methods for efficient DNA extraction from Rice (Oryza sativa L.) (벼 분자육종을 위한 CTAB DNA 추출 시스템 개량)

  • Lee, Jong-Hee;Kwak, Do-Yeon;Yeo, Un-Sang;Kim, Choon-Song;Jeon, Myeong-Gi;Kang, Jong-Rae;Park, Dong-Soo;Shin, Mun-Sik;Oh, Byeong-Geun;Hwang, Hung-goo
    • Korean Journal of Breeding Science
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    • v.40 no.3
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    • pp.286-290
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    • 2008
  • Many important traits have been tagged allowing plant breeders to apply marker assisted selection (MAS) in rice. PCR itself is simple to set up, and requires little hands-on time. However, a crucial limiting step of MAS programs is the reliable and efficient extraction of DNA which can be performed on thousands of individuals. In this study, We describe a modification of the DNA extraction method, in which cetyltrimethylammonium bromide (CTAB) is used to extract DNA from leaf tissues for suitable MAS in rice. We followed the standard 2% CTAB extraction method in all the procedure. In addition we used the 1.2 ml 8-strip tube instead of 1.5 ml E-tubes to fit the 8-multichannel pipette and employ the 96 well plate to use the swing bucket centrifuge. Our modified CTAB DNA extraction method offers several advantages with respect to traditional and simple methods. 1) adult leaf samples collected in paddy field are applicable. 2) 96 leaf samples can be homogenized only one-time by using tungsten carbonate bead and 96well block. 3) semiautomatic loading method using 8-multichannel pipette from DNA extraction to electrophoresis of PCR products. 4) our system can extract about 400 leaf samples per day by only one technicion. Therefore, this method could be useful for marker assisted breeding in rice.

Antioxidant activities and inhibitory effects on oxidative DNA damage of leaf from Zelkova serrata with ethyl acetate fractions and hot water extracts (느티나무 잎 에틸아세테이트 분획물 및 열수 추출물의 항산화 및 산화적 DNA 손상 억제 활성)

  • Jang, Tae-Won;Park, Jae-Ho
    • Journal of Applied Biological Chemistry
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    • v.59 no.3
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    • pp.255-260
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    • 2016
  • Reactive oxygen species (ROS) has been played a critical role in damage of DNA. Recently, many effort is focusing to develop the natural antioxidants for controlling ROS. Zelkova serrata, Ulmaceae, is close as plants which are planted in front of Korea villages. Although Zelkova serrata is familiar with Koreans, those of antioxidant activities and protective effects on oxidative DNA damage haven't studied. We demonstrated antioxidant activities and inhibitory effects on oxidative DNA damage of Leaf from Zelkova serrata with ethyl acetate fractions (EA) and hot water extracts (HW). Between the extracts, EA showed higher activities in 1,1-diphenyl-2-picryl-hydrazyl, 2,2'-azino-bis[3-ethylbenzthiazoline-6-sulphonic acid radical scavenging, $Fe^{2+}$ chelating and reducing power than HW. Also, those of total phenolic content are 56.63 and 51.61 mg/g respectively. In addition, ${\phi}X$-174 RF I plasmid DNA cleavage assay for inhibitory effect by oxidative DNA damage was both EA and HW has significant protective effect on oxidative DNA damage. The results suggested that leaf from Zelkova serrata with ethyl acetate fractions and hot water extracts have surpassing potential as natural resources with antioxidant and inhibitory effect on oxidative DNA damage.

Antioxidant Activity and DNA Damage Protective Effect of a Robinia pseudoacacia L. Flower Extract (아까시 꽃 추출물의 항산화 활성 및 DNA 손상 억제 효과)

  • Kim, Su-Jung;Seo, Go-Un;Seo, Bo-Young;Park, Eun-Ju;Lee, Seung-Cheol
    • Korean journal of food and cookery science
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    • v.27 no.4
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    • pp.99-106
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    • 2011
  • The antioxidant properties of Robinia pseudoacacia L. water and 70% ethanol extracts were evaluated by determining total phenolic content (TPC), DPPH radical scavenging activity (RSA), and reducing power (RP). The water extract showed higher TPC (9.07 mg/g gallic acid equivalents) and RP than those of ethanol extract, whereas ethanol extract had greater DPPH RSA. The R. pseudoacacia L. extracts also showed antigenotoxic effects for 200 ${\mu}M$ $H_2O_2$-induced DNA damage in human leukocytes. The 200 ${\mu}M$ $H_2O_2$-induced DNA damage decreased following treatment with the water extract. Reductions in DNA damage with 50 ${\mu}g/ml$ of the water and ethanol extracts were 46.5 and 32.4%, respectively.

Effect of Acanthopanax extract on the DNA and erythrocyte damage induced by herbicides (제초제로 인한 DNA와 적혈구 손상에 미치는 오가피 추출물의 효과)

  • Seo, Yoo-Na;Kim, Jum-Ji;Sung, Kwang-Soo;Lee, Mi-Young
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.11 no.12
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    • pp.4922-4927
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    • 2010
  • In order to investigate whether the ethanol extract of Acanthopanax sp. might inhibit herbicide-induced DNA damage and erythrocyte damage, the suppression of the oxidative DNA damage of lymphocyte and erythrocyte damage in the presence of the extract were evaluated by comet assay and hemolysis assay, respectively. Phenoxy herbicides, named 2,4-D (2,4-dichlorophenoxyacetic acid) and 2,4,5-T (2,4,5-trichlorophenoxyacetic acid) and bipyridyl herbicide paraquat induced oxidative DNA damages of lymphocytes. However, the oxidative DNA damage by 2,4-D, 2,4,5-T or paraquat was inhibited in vitro upon treating Acanthopanax extract. Moreover, the erythrocyte damage was also suppressed in vitro by Acanthopanax extract treatment.

Protective Effect of Garlic (Allium sativum L.) Extracts Prepared by Different Processing Methods on DNA Damage in Human Leukocytes (마늘의 조리방법에 따른 DNA 손상 보호 효과의 비교)

  • Kim, Jung-Mi;Jeon, Gyeong-Im;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.6
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    • pp.805-812
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    • 2010
  • DNA damage including base modifications, loss of base and breaks in DNA strands can occur by exposure to irradiation, smoking and several components of food. Unrepaired DNA damage is known to lead to cellular dysfunction, cell death, cancer, and other diseases such as arteriosclerosis and diabetes. The protective effect of garlic on oxidative stress induced DNA damage has been reported recently. In this study, we investigated the protective effect of garlic extracts prepared by different processing methods (raw garlic extracts, RGE; grilled garlic extracts, GGE; pickled garlic extracts, PGE) on leukocytic DNA damage using comet assay. Human leukocytes were incubated with ethanol and methanol extract of garlic at various concentrations (1, 5, 10, 50 ${\mu}g$/mL), followed by oxidative stimuli (200 ${\mu}M$ $H_2O_2$ or 200 ${\mu}M$ 4-hydroxynonenal (HNE)). The methanol and ethanol extracts of RGE, GGE, and PGE showed inhibitory activities of DNA damage induced by $H_2O_2$ or HNE. Especially methanol extract of RGE ($ED_{50}$; 13.3 ${\mu}g$/mL) had a higher antigenotoxic effect on $H_2O_2$ induced DNA damage than those of GGE (23.5 ${\mu}g$/mL) or PGE (24.5 ${\mu}g$/mL). HNE induced DNA damage tended to be effectively inhibited by the lower concentration of all garlic extracts. Therefore, garlic might have protective effects against oxidative DNA damage regardless of processing methods (raw, grilled, pickled) which are the general consumed forms of garlic in Korea.