• Title/Summary/Keyword: DNA parts

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A Study on the TCE/PCE Removal Using Biofiltration and the Microbial Communities Variation Using DGGE Method (생물 여과를 이용한 TCE/PCE제거 및 DGGE법을 이용한 관련미생물 군집변화에 관한 연구)

  • Kim, Eung-In;Park, Ok-Hyun;Jung, In-Gyung
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.11
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    • pp.1161-1169
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    • 2008
  • The removals of TCE and PCE vapor with or without a supply of toluene as a primary substrate were compared in a biofiltration process, and the variations of microbial communities associated with the removal were also investigated. As a result of investigations on the removals of TCE/PCE in a biofilter B within which TCE/PCE-acclimated sludge was attached on the surface of media without a supply of primary substrate, and those in another biofilter A where toluene-acclimated sludge was attached with a supply of toluene as a primary substrate, followings were found: (i) parts of microbes responsible to the decomposition of toluene vapor participate in the removal of chlorinated VOCs such as TCE and PCE, and (ii) effective biological removals of TCE and PCE vapor do not necessarily need cometabolism. Sequencing of 16S rDNA obtained from the band profile of DGGE (Denaturating Gradient Gel Electrophoresis), it was confirmed that: (i) uncultured alpha proteobacterium, uncultured Desulfitobacterium, uncultured Rhodobacteraceae bacterium, Cupriavidus necator, and Pseudomonas putida were found to be toluene-decomposing microbes, (ii) alpha proteobacterium HTCC396 is a TCE-removing microbe, (iii) Desulfitobacterium sp. is a PCE-decomposing microbe, and (iv) particularly, uncultured Desulfitobacterium sp. is probably a microbe decomposable not only toluene but also various chlorinated VOC vapor including TCE and PCE.

Morphological Characteristics and Genetic Diversity Using the RAPD Technique in the Arksllell, Scapharca broughtosnii (Schrenck) Sronn Korea and China (한국산 및 중국산 피조개, Scapharca broughtonii(Schrenck)의 형태적 특성과 RAPD 기법을 이용한 유전적 분석)

  • LEE Jeong Mee;PARK Ji-Won;YOO Myung Suk;HONG Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.2
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    • pp.297-304
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    • 1997
  • Morphological and Benetic comparision were studied for the arkshell populations of Korea and China. The shell was collected from commercial arkshell bed in Jinhae Bay, Korea and from an hatchery in Young Seong, China. Shell Parameters, number of radial ribs and random amplified polymorphic DNA (RAPD) band were measured. The two populations had morphological and genetic differences. There was significant morphological difference $(P\leq0.05)$ in the ratio of the longest radial rib to the shell length. Posterior parts of the Chinese arkshell were more elongated than those of the Korean arkshell. Number of radial ribs were $36\~41$ for the Korean arkshell and $39\~43$ for the Chinese arkshell. RAPD markers generated tty each of 19 primers were $2\~6$ bands. Genetic diversity between the two Populations was clear since the genetic similarity was very low, not exceeding 0.29. The genetic similarity among the Korean arkshells $(0.58\~0.40)$ was higher than that of the Chinese arkshells $(0.48\~0.32)$.

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A Novel PAX9 Mutation in a Family with Non-Syndromic Oligodontia (비증후군성 부분 무치증 환자에서 PAX9 유전자 돌연변이)

  • Lee, Ye ji;Shin, Teo Jeon;Hyun, Hong-Keun;Kim, Jung-Wook;Lee, Sang-Hoon;Kim, Young-Jae
    • Journal of the korean academy of Pediatric Dentistry
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    • v.43 no.3
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    • pp.299-305
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    • 2016
  • The aim of this study was to identify the causative genetic mutation in a family with non-syndromic oligodontia. The 7-year-old female proband and her mother underwent oral examination, panoramic radiographs were obtained and blood samples were collected. All exons of the PAX9 gene were amplified by polymerase chain reaction and sequenced. The sequencing results were compared with the standard human gene sequence. The proband lacked 11 permanent teeth, and her mother lacked 19 permanent teeth. No other birth defects were observed. As a result of gene analysis, there was a novel heterozygous nonsense mutation (c.184G>T, $p.Glu62^*$) in exon 2 in both affected subjects. It is suspected that the nonsense mutation leads premature termination of translation, yields a truncated protein 280 amino acids shorter than the wild-type protein. These defects include parts of the paired box domain, a DNA-binding site that plays an essential role in protein function. Otherwise, more likely the mutant transcript would be degraded by nonsense-mediated decay system, resulting haploinsufficiency to cause oligodontia in this family.

Development of Workbench for Analysis and Visualization of Whole Genome Sequence (전유전체(Whole gerlome) 서열 분석과 가시화를 위한 워크벤치 개발)

  • Choe, Jeong-Hyeon;Jin, Hui-Jeong;Kim, Cheol-Min;Jang, Cheol-Hun;Jo, Hwan-Gyu
    • The KIPS Transactions:PartA
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    • v.9A no.3
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    • pp.387-398
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    • 2002
  • As whole genome sequences of many organisms have been revealed by small-scale genome projects, the intensive research on individual genes and their functions has been performed. However on-memory algorithms are inefficient to analysis of whole genome sequences, since the size of individual whole genome is from several million base pairs to hundreds billion base pairs. In order to effectively manipulate the huge sequence data, it is necessary to use the indexed data structure for external memory. In this paper, we introduce a workbench system for analysis and visualization of whole genome sequence using string B-tree that is suitable for analysis of huge data. This system consists of two parts : analysis query part and visualization part. Query system supports various transactions such as sequence search, k-occurrence, and k-mer analysis. Visualization system helps biological scientist to easily understand whole structure and specificity by many kinds of visualization such as whole genome sequence, annotation, CGR (Chaos Game Representation), k-mer, and RWP (Random Walk Plot). One can find the relations among organisms, predict the genes in a genome, and research on the function of junk DNA using our workbench.

An Experimental Study on Apoptosis of Cultivated Wild Ginseng Distilled Herbal Acupuncture by controlled pH and Electrolyte (pH 및 전해질 조절 산양산삼(山養山蔘) 증류약침(蒸溜藥鍼)의 Apoptosis에 관(關)한 실험적(實驗的) 연구(硏究))

  • Chu, Ching sheng;Lee, Sun-gu;Kwon, Ki-rok
    • Journal of Acupuncture Research
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    • v.21 no.6
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    • pp.1-17
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    • 2004
  • Objective : To compare and examine how adjustment of pH and electrolyte can affect the efficacy of cultivated wild ginseng distilled herbal acupuncture, we've administered pure cultivated wild ginseng distilled herbal acupuncture and pH and electrolyte adjusted cultivated wild ginseng distilled herbal acupuncture on A549 human lung cancer lines. Then mRNA and proteins which take parts in apoptosis were examined. Methods : Pure cultivated wild ginseng distilled herbal acupuncture treated group was set as the control group and pH and electrolyte adjusted cultivated wild ginseng distilled herbal acupuncture groups were administered on A549 human lung cancer lines. Cell toxicity was carefully examined and from the analysis of DNA fragmentation, RT-PCR, and Western blot, manifestation of mRNA and proteins which are associated with apoptosis were inspected. Results : The following results were obtained on apoptosis of A549 human lung cancer lines after administering pH and electrolyte adjusted cultivated wild ginseng distilled herbal acupuncture. 1. Measuring cell toxicity of lung cancer cells, higher cell toxicity was detected at pH and electrolyte adjusted groups and the results were concentration dependent. 2. Through DNA fragmentation, we were able to confirm cell destruction in all groups. 3. Experiment groups treated with cultivated wild ginseng distilled herbal acupuncture showed inhibition of Bcl-2 and COX-2 at mRNA and Protein level, whileas increase of Bax was shown. 4. Manifestation of p21, p53, Cyclin E, and Cyclin D1 were confirmed in all groups. 5. Extrication of Cytochrome C was detected at all groups, as well as increased activity of the enzyme caspase-3 and caspase-9, and PARP fragmentation were confirmed. Conclusions : From the above results, we can carefully deduce cell destruction of A549 human lung cancer lines were induced by Apoptosis. At the same concentration level, cell destruction efficacy was better with adjusted pH and electrolyte. Cultivated wild ginseng distilled herbal acupuncture also showed decrease of Bcl-2 and COX-2, as well as increase of Bax. Since cultivated wild ginseng distilled herbal acupuncture increases manifestation of p21, p53, Cyclin E, and Cyclin D1, it affects cellular cycle and through these phenomena, we can consider extrication of Cytochrome C, increase of caspase, and PARP fragmentation are the results.

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Analysis of antigenic domain of GST fused major surface protein (p30) fragments of Toxoplasma gondii (융합단백질로 발현된 톡소포자충의 주요막단백질(p30) 절편의 항원성)

  • 남호우;임경심
    • Parasites, Hosts and Diseases
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    • v.34 no.2
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    • pp.135-142
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    • 1996
  • Antigenic domain of jai or surface protein (p30) of Toxoplosmc Sondii was analyzed after polymerase chain reaction (PCR) of its gene fragments. Hydrophilic or hydrophobic moiety of amino acid sequences were expressed as glutathione S-transferase (G57) fusion proteins. Fragments of p30 gene were as follows: 737, total p30 open reading frame (ORF) ; S28, total ORF excluding N-terminal signal sequence and C-terminal hydrophobic sequence; Al9, N-terminal 2/3 parts of A28; A19, N-terminal 2/3 of S28; P9, C-terminal 2/3 part of S28; Z9. middle 1/3 of S28; and 29, C-terminal 1/3 of S28. respectively. Primer of each fragment was synthesized to include clamp sequence of EcoR I restriction site. PCR amplified DNA was inserted info GST (26 kDa) expression vector, PGEX-47-1 to transform into Escheri,hia coei (.JM105 strain). G57 fusion proteins were expressed with IPTG induction as 63. 54, 45, 45, 35, 36. and 35 kDa proteins measured by SDS-PAGE. Each fusion protein was confirmed with G57 detection kit. Western blot analysis with the serum of a toxoplasmosis patient revealed antigenicity in proteins expressed by T37. S28, and Al9 but not those by Pl8. X9, Y10, and Z9. Antigenicity of p30 seems to be located either in N-terminal 115 part in the presence of middle 1/3 part or in the oligopeptides between margins of the first and second 1/3 parts.

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Radiosensitization Effects of a Zataria multiflora Extract on Human Glioblastoma Cells

  • Aghamohammadi, Azar;Hosseinimehr, Seyed Jalal;Ghasemi, Arash;Azadbakht, Mohammad;Pourfallah, Tayyeb Allahverdi
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7285-7290
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    • 2015
  • Background: Although radiotherapy is one of the most effective strategies in the treatment of cancers, it is associated with short and long term side effects on normal tissues. Zataria multiflora Boiss (Laminacea) (ZM) has several biological properties such as antioxidant and anti-inflammation activities.Here we investigated cell killing effects of a hydroalcoholic Zataria multiflora extract on cell death induced by ionizing radiation in a human glioblastoma cell line (A172) and human non-malignant fibroblasts (HFFF2) in vitro. Materials and Methods: A172 and HFFF2 cells were treated with a hydroalcoholic extract of dried aerial parts of Zataria multiflora at different concentrations (25, 50, 100, 150 and $200{\mu}g/ml$) and then exposed to ionizing radiation (IR). Cell proliferation and DNA fragmentation were evaluated. Thymol content in the extract was analyzed and quantified by HPLC methods. Results: A172 cell proliferation was significantly inhibited by ZM. The percentage cell survival was $91.8{\pm}8.57$ for cells treated with $200{\mu}g/ml$ of ZM extract alone while it was $76.0{\pm}4.27$ and $66.2{\pm}8.42$ for cells treated with ZM and exposed to IR at doses of 3Gy and 6Gy, respectively. Radiation-induced apoptosis in A172 cells was significantly increased following treatment with ZM at doses of $200{\mu}g/ml$. ZM extract did not exhibit any enhanced cell killing effects and apoptosis caused by IR on HFFF2 cells. Conclusions: These data show selective radiosensitization effects of ZM in A172 cells apparently due to increased radiation-induced apoptosis.

Attenuation of ROS Generation by KCNE1 Genes in Cisplatin-treated Auditory Cells

  • Kim, Eun Sook;Park, Sang-Ho;Park, Raekil
    • Korean Journal of Clinical Laboratory Science
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    • v.45 no.3
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    • pp.114-119
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    • 2013
  • Potassium is essential for the proper functioning of the ears. The inner ear's endolymph differs from all other extracellular fluids (in its positive potential) and in the ionic compositions in the various parts of the endolymphatic space. Ion concentration of the endolymph is 150 mM of potassium, which is comparable to the concentrations in other organs. Cisplatin (cis-diamminedichloroplatinum II: CDDP) is one of the most effective anticancer drugs, widely used against various tumors. However, its clinical use is limited by the onset of severe side effects, including ototoxicity and nephrotoxicity. For ototoxicity, a number of evidences in cytotoxic mechanism of cisplatin, including perturbation of redox status, increase in lipid peroxydation, and formation of DNA adduct, have been suggested. Therefore, in this study, the author investigated the relationship between the potassium ions on cisplatin-induced cytotoxicity in HEI-OC1 cells associated with reactive oxygen species (ROS). KCNE1 gene expression by the concentration of intracellular potassium appeared in the plasma membrane and increased the concentration of intracellular potassium. Cisplatin decreased the viability of HEI-OC1 cells, but the KCNE1 gene increased. Also, the KCNE1 gene significantly suppressed generation of intracellular ROS by cisplatin. Western blot analysis showed that the KCNE1 gene increased phase II detoxification enzymes markers such as superoxide dismutase 1 (SOD1), superoxide dismutase (SOD2), NAD(P)H:quinine oxidoreductases (NQO1), which were associated with the scavenger of ROS. These results suggest that the KCNE1 gene for intracellular potassium concentration ultimately prevents ROS generation from cisplatin and further contributes to protect auditory sensory hair cells from ROS produced by cisplatin.

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Allelic variation of melanocortin-1 receptor locus in Saudi indigenous sheep exhibiting different color coats

  • Mahmoud, Ahmed H.;Mashaly, Ashraf M.;Rady, Ahmed M.;Al-Anazi, Khalid M.;Saleh, Amgad A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.2
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    • pp.154-159
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    • 2017
  • Objective: This study was designed to characterize the DNA polymorphisms of the melanocortin-1 receptor (MC1R) gene in indigenous Saudi Arabian sheep breeds exhibiting different color coats, along with individuals of the Sawaknee breed, an exotic sheep imported from Sudan. Methods: The complete coding region of MC1R gene including parts of 3' and 5' untranslated regions was amplified and sequenced from three the indigenous Saudi sheep; Najdi (generally black, n = 41), Naeimi (generally white with brown faces, n = 36) and Herri (generally white, n = 18), in addition to 13 Sawaknee sheep. Results: Five single nucleotide polymorphisms (SNPs) were detected in the MC1R gene: two led to nonsynonymous mutations (c.218 T>A, p.73 Met>Lys and c.361 G>A, p.121 Asp>Asn) and three led to synonymous mutations (c.429 C>T, p.143 Tyr>Tyr; c.600 T>G, p.200 Leu>Leu, and c.735 C>T, p.245 Ile>Ile). Based on these five SNPs, eight haplotypes representing MC1R $E^d$ and $E^+$ alleles were identified among the studied sheep breeds. The most common haplotype (H3) of the dominant $E^d$ allele was associated with either black or brown coat color in Najdi and Sawaknee sheep, respectively. Two other haplotypes (H6 and H7) of $E^d$ allele, with only the nonsynonymous mutation A218T, were detected for the first time in Saudi indigenous sheep. Conclusion: In addition to investigating the MC1R allelic variation in Saudi indigenous sheep populations, the present study supports the assumption that the two independent nonsynonymous Met73Lys and Asp121Asn mutations in MC1R gene are associated with black or red coat colors in sheep breeds.

Expression of Luteinizing Hormone (LH) Subunit Genes in Mouse Testis

  • Kim, Hee Soo;Lee, Sung-Ho
    • Development and Reproduction
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    • v.21 no.3
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    • pp.327-333
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    • 2017
  • Gonadotropins are heterodimers consisting an alpha chain ($Cg{\alpha}$) and a beta chain. Interestingly, presence of complicated $LH-{\beta}$ transcripts in rat testis was accidently found; testicular $LH-{\beta}$ transcripts were confined in seminiferous tubules to spermatids, and the translated products were localized in the elongated spermatids. We hypothesized that mouse testis has potential to produce the tissue specific $LH-{\beta}$ with similar structure to the rat testicular forms. To verify our hypothesis, we examined the adult mouse (ICR) testis using RT-PCR and immunohistochemistry. The PCR revealed the presence of the identical products in the reactions for three LH subunit types. The expected product sizes for mouse $Cg{\alpha}$ and $LH-{\beta}$ known as pituitary type were 224 bp and 503 bp, respectively. The testicular type $LH-{\beta}$ products were produced by a primer set based on the rat sequences, with unexpected size of 800 bp. Sequencing revealed that the proximal and distal parts (2-82 and 661- 773 bp, respectively) were homologous to rat testicular $LH-{\beta}$ cDNA, and middle part (83-660 bp) was a unique mouse-specific region. Both $Cg{\alpha}$ and $LH-{\beta}$ positive signals were in the round and elongated spermatids and mature sperms, and the $LH-{\beta}$ signals were more intense. In conclusion, our study demonstrated that the presence and localization of the LH subunits in mouse testis. Further studies will be needed to understand the precise structure and function of mouse testicular LH.