• Title/Summary/Keyword: DNA회복

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Chaperon Effects of Campylobacter jejuni groEL Genes Products in Escherichia coli (Campylobacter jejuni의 groEL 유전자 산물의 대장균에서의 Chaperon효과)

  • Lim, Chae-Il;Kim, Chi-Kyung;Lee, Jae-Kil
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.47-52
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    • 1994
  • The cells of Campylobacter jejuni heat-shocked at 48${\circ}C$ for 30 min synthesized the heat shock proteins of HSP90, HSP66 and HSP60. Those heat shock proteins were found to correspond to the heat shock proteins of HSP87, HSP66 (DnaK), and HSP58 (GroEL) of E. coli, respectively. By Southern blot analysis of the chromosomal DNAs of C. jejuni with groESL and dnaK genes of E. coli as DNA probes, the heat shock genes of C. jejuni which are homologous to the E. coli groESL and dnaK genes were found to exist in the chromosomal DNA. The genomic libraries of C. jejuni were constructed with the cosmid vector pWE15 and the groEL gene of C. jejuni were cloned in E. coli B178 groEL44 temperature senstive mutant. The hybrid plasmid (pLC1) was inserted with the DNA fragment (about 5.7kb in size) containing the groEL gene. E. coli groEL44 mutant cell transformed with the pLC1 could grow at 42${\circ}C$ by synthesizing the HSP60 of C. jejuni and regained the susceptibility to the ${\lambda}$ vir phage by expression of the groEL gene in the cloned cells. These indicated that the groEL products of C. jejuni had chaperon effects by synthesizing the heat shock proteins in the cloned cells of E. coli.

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Quantification of triterpenes in Centella asiatica cultivated in a smart farm, and their effect on keratinocyte activation (스마트팜 재배 병풀의 triterpenes 정량 및 각질형성세포 활성화 효과)

  • Jin Hong Park;Seong Min Jo;Da Hee Lee;Youngmin Park;Hwan Bong Chang;Tae Jin Kang;Kiman Lee
    • Food Science and Preservation
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    • v.30 no.3
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    • pp.483-491
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    • 2023
  • This study aimed to compare the bioactive compounds in Centella asiatica (C. asiatica) cultivated in a smart farm and a field and their effects on human keratinocyte cells. C. asiatica was collected in Jeju-do, Korea, and cultured in a smart farm and a field. The main bioactive compounds in the two differentially cultured C. asiatica were identified, and their activation in keratinocytes were assessed. Amplification and sequencing of the internal transcribed spacer (ITS) DNA in the nucleus and psbA-H DNA in the chloroplast were performed for species analysis. A comparison of DNA of plants reported in the NCBI GenBank was performed. The ITS DNA and psbA-H DNA sequences of C. asiatica cultivated in a smart farm and a field were consistent with No. MH768338.1 and No. JQ425422.1, respectively. Analysis of the triterpenes was performed using high performance liquid chromatography (HPLC) and as a result, C. asiatica cultured in a smart farm had more triterpenes than those cultured in a field. The effects of C. asiatica grown in a smart farm on cell proliferation and scratch recovery in HaCaT cells were greater than those grown in a field. These results suggest that C. asiatica cultivated in a smart farm can be effectively utilized as a health functional food.

Bacillus sphaericus 의 ts-D1216의 특성연구(I)

  • 김재수;서정희;황성희;이형환
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1986.12a
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    • pp.521.1-521
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    • 1986
  • B. sphaericus의 sporeless ts-D1216 돌연변이체의 유전학적 특성을 방사성 동위원소를 이용하여 DNA의 합성과 RNA의 합성을 측정하였고 일반적 특성을 연구하였다. ts-D1216 돌연변이체의 대수증식기 세포를 3$0^{\circ}C$에서 제한온도 (42$^{\circ}C$)로 이동시켰을 때 RNA의 합성은 4-5시간 정상적으로 합성이 계속 일어났고, DNA의 합성은 60-100분까지는 정상적인 율로 일어나다가 그 후에 결정적으로 감소되었다. 또한, DNA합성이 멈춘후에도 세포수는 증가했고, 4$0^{\circ}C$에서 성장기간이 더 길면 길수록 DNA합성에의 회복능력이 상실되었다.

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Protective Effect of Electrolyzed Reduced Water on the Paraquat-induced Oxidative Damage of Human Lymphocyte DNA (Paraquat에 의한 사람 임파구 DNA 손상에 대한 환원전리수의 보호효과)

  • Park, Eun-Ju;Ryoo, Kun-Kul;Lee, Yoon-Bae;Lee, Jong-Kwon;Lee, Mi-Young
    • Applied Biological Chemistry
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    • v.48 no.2
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    • pp.155-160
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    • 2005
  • Electrolyzed reduced water (ERW), showing extremely negative oxidation-reduction potential, was used to investigate the effects of paraquat-induced damages on DNA from human lymphocyte. The effect of ERW on paraquat-induced oxidative DNA damage in human lymphocytes was evaluated by Comet assay (single-cell gel electrophoresis) quantified as percentage fluorescence in tail. Comet assay has been used widely to assess the level of the DNA damage in individual cells. Lymphocytes were oxidatively challenged with various concentrations of paraquat for 30 min at $37^{\circ}C$, and were then treated with electrolyzed reduced water for 30 min. The oxidative DNA damage by paraquat, as indicated by the fluorescent tail in DNA, increased in a dose-dependent manner. However, oxidative damage of the DNA was almost completely prevented upon treatment with electrolyzed reduced water.

Effect of DPBll Gene for the Transcriptional Induction by DNA Damage During Cell Cycle in Saccharomyces cerevisiae (출아효모의 세포주기동안 DNA 상해에 의한 발현 유도에 미치는 DPB11 유전자의 영향)

  • 선우양일;임선희;배호정;김중현;김은아;김승일;김수현;박정은;김재우
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.96-102
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    • 2002
  • The S-phase checkpoint mechanisms response to DNA damage or inhibition of DNA replication for maintenance of genetic stability in eukaryotic cells. These roles include cell cycle control arrest at S-phase and Iranscriptional induction of repair genes. To characterize the defects of dpbll mutant for both these responses, we examined the over-expression effect of DPBll gene, the sensitivity to HU, MMS, and the transcriptional pattern by DNA damage agent for RNRS mRNA. RNRS transcript is induced in response to a wide variety of agents that either damage D7A directly through chemical modification or induce stress by blocking DNA synthesis. As results, dpbll-1 cells are sensitive to DNA damage agents and the level of RNR3 mRNA is reduced approximately 40% than wild type cells. Moreover, we found the same results in dpb2-1 cells. Therefore, we propose that DPB2 and DPBll act as a sensor of replication that coordinates the transcriptional and cell cycle responses to replication blocks.

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.

DNA damages with Fpg/Endo Ⅲ FLARE Assay in cynomolgus monkeys exposed to stainless steel welding fume (용접흄 흡입노출 영장류에서 Fpg/Endo Ⅲ FLARE Assay를 이용한 DNA 손상 및 회복)

  • Rim, Kyung Taek;Kim, Soo Jin;Chung, Yong Hyun;Kim, Hyeon Yeong;Maeng, Seung Hee;Yu, Il Je
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.17 no.4
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    • pp.272-281
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    • 2007
  • 선박제조업을 비롯한 운송업 및 건축업 등의 다양한 분야에서 용접기술이 이용되어 옴에 따라 용접근로자들에 대한 산업보건학적 관심이 높아지고 있다. 노출정도가 다양하기는 하지만 용접흄은 6가 크롬을 비롯한 금속화합물과 유해가스, 화학물질 등을 복합적으로 포함하고 있는 스테인레스 스틸 용접흄에 대한 유전독성영향을 평가하기 위하여 흡입챔버를 이용, 실험동물인 영장류에 스테인레스 스틸 용접흄을 노출시키고 혈액 내 lymphocytes에 생성된 용접흄 노출농도 및 시간별 DNA 손상정도 및 그 회복효소를 측정함으로써, 유해성이 완전하게 확인되지 않은 용접흄에 노출되어 나타날 수 있는 암을 비롯한 심각한 건강영향을 예방하기 위한 각 지표들을 찾아 그 유용성을 비교하고자 하였다. 영장류를 노출시키기 위해 robotic arm을 장치한 영장류 흡입노출 시스템을 개발하였으며, 이 노출 시스템을 이용하여 수컷 영장류 6마리에 대해 용접흄 노출시험을 실시하였는데 실험군은 대조군 2, 저농도 ($31mg/m^3$) 노출군 2, 고농도 ($63mg/m^3$) 노출군 2마리로 구성하였고, 1일 2시간씩 일주일에 5일 동안 용접흄에 노출시켰다. 노출 농도는 지속적으로 모니터링 하였고, 노출과정 중에 영장류의 혈액을 채취하여 lymphocytes를 분리, 단세포 DNA 손상을 선별하기 위해 DNA 손상회복 효소인 E. coli formamidopyrimidine-DNA glycosylase (Fpg)와 endonuclease Ⅲ (Thymine Glycol-DNA glycosylase) 투여와 Comet asaay (single cell gel electrophoresis, 단세포겔전기영동기법)를 결합시켜 이용하는 Fpg/Endo III FLARE 분석법을 사용하였다. Fpg enzyme에 의한 olive tail moment값의 변화는 16주 노출군부터 노출부검(34주)군 까지 노출농도가 높아짐에 따른 olive tail moment 기하평균 값의 양 반응관계를 보기는 어렵지만, 고농도군의 경우 27주 노출군에서 가장 높은 olive tail moment 값을 보이고 이후 차츰 감소하였다. 한편 16주에서 22주까지의 노출기간에서는 대조군에 비해 노출군에서 DNA손상정도(olive tail moment값)는 모두 유의하게 높았으나, 6, 12, 18, 25, 31, 33, 35주간 노출하였을 때는 다른 결과를 보였다. 각 실험군의 Fpg enzyme에 의한 tail length값의 분포를 살펴볼 때, 저농도군 및 고농도군에서 27주간 노출하였을 때 가장 높은 tail length 값을 보이고 이후 차츰 감소하는 경향을 보였다. 또한 16, 22주간 노출하였을 때 대조군에 비해 노출군에서 tail length 값이 유의하게 높았으나, 20주간에서만 양 반응관계가 관찰되었고, 다른 주간에서는 양 반응 및 기간 반응관계를 나타내지는 않았다. Endo III enzyme에 의한 olive tail moment값의 변화는 기간별 노출군에서 대조군에 비해 높은 DNA손상정도(olive tail moment값)를 나타내는 결과들이 있었지만, 10, 12, 16, 22, 25, 31주간 노출하였을 때 등 상당수 노출기간에서 반응관계를 나타내지는 않았다. 각 실험군의 Endo III enzyme에 의한 tail length값의 분포를 살펴볼 때, 18, 20, 27, 33주간 노출하였을 때 대조군에 비해 노출군에서 tail length 값이 조금 높았지만, 양 반응 및 기간 반응관계를 보이지 않았고 수치의 크기가 불규칙하게 변화하였다. 즉, DNA에 있어 산화된 pyrimidine을 형성하여 손상된 부위의 염기를 제거함으로써 AP site (abasic site)를 만들고 이들이 Comet assay를 통해 break로 전환된 것을 포함한 DNA손상을 측정하기 위하여 endonuclease III (Endo III)를 첨가시킨 Endo III FLARE 분석법을 실시한 결과, 본 연구에서 나타난 결과는 용접흄 노출 영장류에서 Olive tail moment 및 tail length 공히 노출량 및 노출기간 반응관계를 볼 수 없었다. Endo III FLARE 분석법을 통한 산화적 DNA 손상지표는 영장류에 적용하기에는 적응반응현상으로 대조군과 유의한 차이도 관찰할 수 없었고 더욱이 역으로 대조군에서의 자연발생적 수치가 더 높아질 수 있어 용접흄 노출 영장류의 모니터링 지표로 사용하기에는 제한점이 있었다.

Mutator effects of plasmid pKM101 and pSL4 to E. coli DNA repair (E. coli DNA 회복에 미치는 플라스미드 pKM101과 pSL4의 mutator 기능)

  • 전홍기;이상률;백형석
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.109-113
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    • 1990
  • The mutagenesis-enhancing plasmid pKM101 and its mutant pSL4 were introduced into Escherichia coli B/r strains possessing different DNA repair capacities ($phr^{-}, recA^{-}, uvrA^{-}, uvrB^{-}$) and determined the protection effect and mutagenecity for UV and MNNG. The mutability and protection effect of plasmid pKM101 and pSL4 were affected by different DNA repair capacity. The mutagenecity and resistance of two plasmids were increased against UV and MNNG, and plasmid pSL4 had a higher effect than pKM101. We suggest that the functional differences between pKM101 and pSL4 is due to the variety of mutator gene.

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Regulatory Expression of DNA Repair Genes Involved in Adaptive Response (적응반응 관련 DNA 회복유전자의 발현조절에 관한 연구)

  • 최수영;이희원;박상대
    • Environmental Mutagens and Carcinogens
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    • v.10 no.1
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    • pp.1-10
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    • 1990
  • The regulation of DNA repair genes expression was investigated using fused genes, in which the promoter of repair genes was hybridized with the lacZ structural gene. The activities of beta-galactosidase expressed from the fused gense were highly increased when the host cells were exposed to methylating agents, such as methyl methansulfonate (MMS), N-methyl-N'-nitro-nitrosoguanidine (MNNG) and methyl nitrosourea (MNU). On the other hand, the enzyme activities from the fused genes were not induced when the cells were treated with ethylating or nonalkylating agents, such as ethyl methansulfonate (EMS), 4-nitroquinoline-1-oxide (4NQO), Bleomycin, and Benzo(a)pyrene (BP).

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Effect of trichostatin A on NF-κB DNA binding activity and myogenesis in C2Cl2 skeletal muscle Precursor cell (C2C12 근육아세포에서 trichostatin A에 의한 NF-κB DNA 결합 활성과 근육발생에 미치는 영향)

  • 임운기;김경창;신혜자
    • Journal of Life Science
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    • v.12 no.1
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    • pp.55-60
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    • 2002
  • The differentiation of skeletal muscle precursor cells in culture is marked by the transcriptional activation of muscle-specific genes and the morphological differentiation of myoblast into multinucleate myotube. In this study, we examined the effect of TSA (Trichostatin A) on WF-kB DNA binding activity and muscle cell fusion in the process of myogenesis. Under TSA treatment, C2C12 myoblast could not fuse to myotube and its NF-kB DNA binding activity was also blocked. To investigate whether these phenomenons were affected by TSA in direct or not, differentiation media (DM) used to differentiate cells without TSA was concentrated and added to C2C12 myoblast with TSA simultaneously. C2C12 myoblast was fused to myotube and NF-kB DNA binding activity was recovered. These results suggest that TSA affects on the differentiation of myoblast, perhaps through several factors, by inhibiting myoblst fusion and blocking NF-kB DNA binding activity.