• Title/Summary/Keyword: Single strand DNA

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Phage Particle Proteins and Genomic Characterization of the Lactobacillus plantarum Bacteriophage SC 921. (Lactobacillus plantarum Bacteriophage SC 921의 phage particle protein 및 genome의 특성)

  • 김재원;신영재;심영섭;유승구;윤성식
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.117-121
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    • 1998
  • Bacteriophage SC 921 of Lactobacillus plantarum, isolated from kimchi, showed high lytic effects at 0.2 M.O.I. level. The phage particle contained 4 major proteins (48, 34, 32, 29 kDa). Intact DNA of phage SC 921 is a double stranded linear molecule, and the genomic size is approximately 66.5 kilobase pairs (kbp). Restriction analysis of the genome showed that Sma I gave single site cut and Xba I gave 2 site cuts, while Cla I, Kpn I, and EcoR I formed 4, 5, and 6 cuts, respectively. Hind III digested phage DNA to many fragments. A restriction map of genomic DNA was constructed using the restriction endonuclease Kpn I, Sma I, and Xba I. Bacteriophage SC 921 was compared with B2 phage which had been reported to infect Lactobacillus plantarum ATCC 8014(KCCM l1322). Bacteriophage SC 921 differs from B2 phage at least in thr size of its genome and phage particle proteins.

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Genotoxicity and Identification of Differentially Expressed Genes of Formaldehyde in human Jurkat Cells

  • Kim, Youn-Jung;Kim, Mi-Soon;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.1 no.4
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    • pp.230-236
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    • 2005
  • Formaldehyde is a common environmental contaminant found in tobacco smoke, paint, garments, diesel and exhaust, and medical and industrial products. Formaldehyde has been considered to be potentially carcinogenic, making it a subject of major environmental concern. However, only a little information on the mechanism of immunological sensitization and asthma by this compound has been known. So, we performed with Jurkat cell line, a human T lymphocyte, to assess the induction of DNA damage and to identify the DEGs related to immune response or toxicity by formaldehyde. In this study, we investigated the induction of DNA single strand breaks by formaldehyde using single cell gel electrophoresis assay (comet assay). And we compared gene expression between control and formaldehyde treatment to identify genes that are specifically or predominantly expressed by employing annealing control primer (ACP)-based $GeneFishing^{TM}$ method. The cytotoxicity ($IC_{30}$) of formaldehyde was determined above the 0.65 mM in Jurkat cell in 48 h treatment. Based on the $IC_{30}$ value from cytotoxicity test, we performed the comet assay in this concentration. From these results, 0.65 mM of formaldehyde was not revealed significant DNA damages in the absence of S-9 metabolic activation system. And the one differentially expressed gene (DEG) of formaldehyde was identified to zinc finger protein 292 using $GeneFishing^{TM}$ method. Through further investigation, we will identify more meaningful and useful DEGs on formaldehyde, and then can get the information on the associated mechanism and pathway with immune response or other toxicity by formaldehyde exposure.

Characterization of Single Stranded DNA-Dependent ATPase Activities of Deinococcus radiodurans RecA Protein (Deinococcus radiodurans RecA 단백질의 외가닥 DNA-의존성 ATPase 활성 분석)

  • Kim, Jong-Il
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.250-255
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    • 2007
  • The RecA protein of Deinococcus radiodurans is essential for the extreme radiation resistance of this organism. The central steps involved in recombinational DNA repair require DNA-dependent ATP hydrolysis by recA protein. Key feature of RecA protein-mediated activities is the interactions with ssDNA and dsDNA. The ssDNA is the site where RecA protein filament formation nucleates and where initiation of DNA strand exchange takes place. The effect of sequence heterogeneity of ssDNA was examined in this experiment. The rate of homopolymeric synthetic ssDNA-dependent ATP hydrolysis was constant or nearly so over a broader range of pHs. For poly(dT)-dependent ATP or dATP hydrolysis, rates were generally faster, with a broader optimum between pH 7.0 and 8.0. Activities of RecA protein were affected by the ionic environment. The ATPase activity was shown to have different sensitivity to anionic species. The presence of glutamate seemed to slimulate the hydrolytic activity. Dr RecA protein was shown to require $Mg^{2+}$ ion greater than 2 mM for binding to etheno ssDNA and the binding stoichiometry of 3 nucleotide for RecA protein monomer.

Antimicrobial and Antioxidative Effects of Roselle (Hibiscus sabdariffa L.) Flower Extract and Its Fractions on Skin Microorganisms and Oxidation

  • Kang, Pil-Sung;Seok, Jae-Hwan;Kim, Yo-Han;Eun, Jae-Soon;Oh, Suk-Heung
    • Food Science and Biotechnology
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    • v.16 no.3
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    • pp.409-414
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    • 2007
  • Antimicrobial and antioxidative effects of 14 different herbal flower extracts on skin microorganisms and oxidation were tested in this research. Herbal flower extracts were prepared with 70% ethanol. Among the herbal flower extracts, roselle (Hibiscus sabdariffa L.) flower extract showed the highest antimicrobial activity against Staphylococcus epidermidis as determined by a paper disc method. The seventy % ethanol extract of roselle flower was fractionated by sequential hexane, chloroform, ethyl acetate, n-butanol, and water fractionation. The growth of S. epidermidis, Streptomyces collinus, Streptomyces coeruleoprunus, Salmonella enteritidis, Vibrio parahaemolyticus, and Malassezia pachydermatis was most efficiently inhibited by ethyl acetate fraction of roselle flower extract as determined by a paper disc method and growth inhibition curves. In addition, the ethyl acetate fraction, water fraction and butanol fraction showed free radical scavenging and DNA cleavage inhibition activities. These results demonstrate that roselle flowers hold antimicrobial and antioxidative activities against skin microorganisms and oxidants.

Effect Evaluation of Benzo[a]pyrene on Multiple Biomarkers in Common Carp (Cyprinus carpio) (잉어 (Cyprinus carpio)의 다중바이오마커를 이용한 Benzo[a]pyrene의 영향평가)

  • Kim, Woo-Keun;Kim, Ja-Hyun;Yeom, Dong-Hyuk;Lee, Sung-Kyu
    • Environmental Analysis Health and Toxicology
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    • v.23 no.3
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    • pp.171-178
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    • 2008
  • 수백여 종의 개별물질이 불완전 연소 혹은 유기물의 열분해로 인해 발생되는 다환방향족 탄화수소(PAHs)는 환경에서 중요한 오염원이 되고 있다. 본 연구는 다양한 바이오마커를 이용하여 수서생태계에 벤조피렌(benzo[a]pyrene)과 같은 다환방향족 탄화수소의 영향을 분석하였고, 이에 대한 통합적 결과 모델을 도출하였다. 즉, 잉어(Cyprinus carpio)를 이용하여 여러 농도의 벤조피렌(3, 12, $34{\mu}g/L$, 측정농도 기준)에 10일간 노출시킨 다음, DNA single-strand break, ethoxyresorufin-O-deethylase (EROD), acetylcholine esterase (AChE)와 vitellogenin (VTG)의 농도를 측정하였다. 벤조피렌은 잉어의 DNA 손상을 유도하였고, 낮은 농도에서 EROD와 VTC의 유의적인 활성을 보였으나, 신경전달물질과 관련이 깊은 AChE 효소활성에는 영향을 미치지 않았다. 이 결과를 star plot를 이용하여 통합 및 분석하였으며, 노출농도에 따른 통합 반응지수(integrated biomarker response value: IBR)로 나타내었다. 이런 다양한 바이오마커의 결과들은 벤조피렌에 대한 어류의 영향과 수생태 모니터링 자료로 이용 가능할 것으로 여겨지며, 통합반응지수는 생태위해성평가에서 유용한 도구로 쓰일 가치가 있는 것으로 평가된다.

Effects of Genotypes on In Vitro Maturation and Fertilization of Frozen-Thawed Porcine Oocytes

  • Jia Y. H.;Jin H. J.;Wee M. S.;Cheong H. T.;Yang B. K.;Park C. K.
    • Reproductive and Developmental Biology
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    • v.29 no.4
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    • pp.207-212
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    • 2005
  • In the present study, we investigated the effects of genotypes on in vitro maturation and fertilization in porcine fresh/frozen-thawed oocytes. The porcine cumulus-oocyte complexes (COCs) were divided into four groups according to whether they were: (1) in vitro matured; (2) cryopreserved and in vitro matured; (3) in vitro fertilized and (4) cryopreserved, and in vitro fertilized. Maturation of porcine COCs was accomplished by incubation in NCSU23 medium. Immature oocytes were cryopreserved by Open Pulled Straws (OPS) method according to Vajta et al., (1998). Oocytes stained by Acetic-Orcein method were observed under the microscope. DNA extracted from the ovaries was analyzed by RAPD (random amplified polymorphic DNA) and SSCP (single strand conformational polymorphisrrt) method. The rates of oocytes maturation and fertilization were significantly high in AA genotype. The results indicated that in vitro maturation and fertilization in porcine fresh/frozen-thawed oocytes may be affected by genotypes in pigs.

THE EFFECT OF GENETIC VARIATION IN THE DNA BASE REPAIR GENES ON THE RISK OF HEAD AND NECK CANCER (DNA 염기손상 치유유전자의 변이와 두경부암 발생 위험성)

  • Oh, Jung-Hwan;Yoon, Byung-Wook;Choi, Byung-Jun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.34 no.5
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    • pp.509-517
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    • 2008
  • DNA damage accumulates in cells as a result of exposure to exogenous agents such as benzopyrene, cigarette smoke, ultraviolet light, X-ray, and endogenous chemicals including reactive oxygen species produced from normal metabolic byproducts. DNA damage can also occur during aberrant DNA processing reactions such as DNA replication, recombination, and repair. The major of DNA damage affects the primary structure of the double helix; that is, the bases are chemically modified. These modification can disrupt the molecules'regular helical structure by introducing non-native chemical bonds or bulky adducts that do not fit in the standard double helix. DNA repair genes and proteins scan the global genome to detect and remove DNA damage and damage to single nucleotides. Direct reversal of DNA damage, base excision repair, double strand break. DNA repair are known relevant DNA repair mechanisms. Four different mechanisms are distinguished within excision repair: direct reversal, base excision repair, nucleotide excision repair, and mismatch repair. Genetic variation in DNA repair genes can modulate DNA repair capacity and alter cancer risk. The instability of a cell to properly regulate its proliferation in the presence of DNA damage increase risk of gene mutation and carcinogenesis. This article aimed to review mechanism of excision repair and to understand the relationship between genetic variation of excision repair genes and head and neck cancer.

한우 불포화지방산 생합성 효소(SCD) 유전자가 도체 및 육질형질에 미치는 영향

  • Sin, Seong-Cheol;Kim, Hui-Chan;Kim, Gi-Rak;Jeong, Hwa-Cheol;Choe, Eun-Ju;Jo, Ha-Na;Jeon, Sang-Hui;Gwon, Su-Yeon;Kim, Bo-Hyeon;Jeong, Gu-Yong;Jeong, Ui-Ryong
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2005.10a
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    • pp.123-126
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    • 2005
  • 본 연구는 고등동물의 불포화지방산 생합성의 핵심 효소로 알려져 있는 Stearoyl-CoA desaturase(SCD) 유전자의 특정한 단일염기다형(single nucleotide polymorphism; SNP)이 한우의 도체 및 육질형질에 미치는 영향을 분석하기 위해 SCD 유전자의 Intron 7번 영역의 특정부위를 포함하는 primer를 제작하고, 염기서열 분석을 통하여 유전자 구조를 해석한 결과 총 211bp 크기를 갖는 염기서열의 122번째에서 아데닌(A)${\leftrightarrow}$구아닌(G) 염기치환으로 발생한 단일염기다형(SNP) 부위를 발견하였다. 이들 단일염기다형 염기서열 부위를 PCR-SSCP(single-strand conformation polymorphism) 기법을 이용하여 분석한 결과 3종류의 SNP 유전자형(A/A, A/G 및 G/G)을 검출하였다. 이 가운데 A/G 유전자형이 한우의 근내지방도와 등지방두께와 고도의 유의적 연관성이 있다는 새로운 사실을 발견하였다. 따라서, 본 연구를 통해 개발된 한우 SCD 유전자의 특정한 단일 염기다형 표지인자는 한우의 연령 및 성별에 관계없이 육질이 우수한 고급육을 생산하는 우량 한우의 조기식별에 매우 유용한 DNA 표지인자로 활용할 수 있을 것으로 기대된다.

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Polymerase chain reaction for the detection of Newcastle disease virus (닭 뉴캐슬병 바이러스의 특이 검출을 위한 polymerase chain reaction 법)

  • Yeo, Sang-geon;Kim, Do-kyoung;Park, Seon-ja
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.565-573
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    • 1998
  • To study the specific tools for the diagnosis of Newcastle disease virus (NDV) in chicken, polymerase chain reaction (PCR) and its presumable conditions were evaluated for the detection of hemagglutinin-neuraminidase (HN) gene of NDV RNA. For these purposes, Kyojeongwon strain of the NDV was propagated in allantoic cavity of SPF embryonating chicken eggs, and viral RNA was extracted from fractionated virus after the allantoic fluids were ultracentrifuged with sucrose gradient. The first-strand cDNA was then made for the HN gene of NDV RNA by reverse transcription at $42^{\circ}C$ for 1 hour using specific primer complementary to the HN gene. The single-stranded cDNA was used as template in the PCR of the HN-DNA, and various conditions of the PCR were evaluated to set up method for the specific detection of the HN-DNA. The PCR conditions promising for the detection of HN gene consist of preheating at $94^{\circ}C$, 5 min, 30 cycles of denaturation at $94^{\circ}C$, 1 min, annealing at $55^{\circ}C$, 1 min and polymerization at $72^{\circ}C$, 2 min, and a cycle of extension at $72^{\circ}C$, 5 min. when NDVs of allantoic fluids without fractionation were applied to the above PCR condition, the HN genes were detected effectively not only from Kyojeongwon but from other velogenic strains such as Herts and a field isolate.

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