• Title/Summary/Keyword: Fragment method

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A DNA Sequence Alignment Algorithm Using Quality Information and a Fuzzy Inference Method (품질 정보와 퍼지 추론 기법을 이용한 DNA 염기 서열 배치 알고리즘)

  • Kim, Kwang-Baek
    • Journal of Intelligence and Information Systems
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    • v.13 no.2
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    • pp.55-68
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    • 2007
  • DNA sequence alignment algorithms in computational molecular biology have been improved by diverse methods. In this paper, we proposed a DNA sequence alignment algorithm utilizing quality information and a fuzzy inference method utilizing characteristics of DNA sequence fragments and a fuzzy logic system in order to improve conventional DNA sequence alignment methods using DNA sequence quality information. In conventional algorithms, DNA sequence alignment scores were calculated by the global sequence alignment algorithm proposed by Needleman-Wunsch applying quality information of each DNA fragment. However, there may be errors in the process for calculating DNA sequence alignment scores in case of low quality of DNA fragment tips, because overall DNA sequence quality information are used. In the proposed method, exact DNA sequence alignment can be achieved in spite of low quality of DNA fragment tips by improvement of conventional algorithms using quality information. And also, mapping score parameters used to calculate DNA sequence alignment scores, are dynamically adjusted by the fuzzy logic system utilizing lengths of DNA fragments and frequencies of low quality DNA bases in the fragments. From the experiments by applying real genome data of NCBI (National Center for Biotechnology Information), we could see that the proposed method was more efficient than conventional algorithms using quality information in DNA sequence alignment.

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A Green's-Matrix Approach to Chemisorption

  • Jang, Yun-Hee;Kim, Ho-Jing
    • Bulletin of the Korean Chemical Society
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    • v.14 no.2
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    • pp.238-243
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    • 1993
  • A self-consistent-field Green's matrix method for the calculation of electronic properties of chemisorbed system is devised and applied to the methanol on copper(110) surface. The method is based on CNDO Hartree-Fock approximation. Contour integration in the complex energy plane is used for an efficient calculation of the charge-density bond-order matrix. The information on each fragment prior to chemisorption is efficiently used and a small number of iterations are needed to reach the self-consistency. The changes of density of states and other quantities of methanol due to chemisorption are consistent with reported experimental results.

Modified T-RFLP Methods for Taxonomic Interpretation of T-RF

  • Lee, Hyun-Kyung;Kim, Hye-Ryoung;Mengoni, Alessio;Lee, Dong-Hun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.624-630
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    • 2008
  • Terminal restriction fragment length polymorphism (T-RFLP) is a method that has been frequently used to survey the microbial diversity of environmental samples and to monitor changes in microbial communities. T-RFLP is a highly sensitive and reproducible procedure that combines a PCR with a labeled primer, restriction digestion of the amplified DNA, and separation of the terminal restriction fragment (T-RF). The reliable identification of T-RF requires the information of nucleotide sequences as well as the size of T-RF. However, it is difficult to obtain the information of nucleotide sequences because the T-RFs are fragmented and lack a priming site of 3'-end for efficient cloning and sequence analysis. Here, we improved on the T-RFLP method in order to analyze the nucleotide sequences of the distinct T-RFs. The first method is to selectively amplify the portion of T-RF ligated with specific oligonucleotide adapters. In the second method, the termini of T-RFs were tailed with deoxynucleotides using terminal deoxynucleotidyl transferase (TdT) and amplified by a second round of PCR. The major T-RFs generated from reference strains and from T-RFLP profiles of activated sludge samples were efficiently isolated and identified by using two modified T-RFLP methods. These methods are less time consuming and labor-intensive when compared with other methods. The T-RFLP method using TdT has the advantages of being a simple process and having no limit of restriction enzymes. Our results suggest that these methods could be useful tools for the taxonomic interpretation of T-RFs.

Open versus closed treatment for extracapsular fracture of the mandibular condyle

  • Lee, Junyeong;Jung, Hee-Yeoung;Ryu, Jaeyoung;Jung, Seunggon;Kook, Min-Suk;Park, Hong-Ju;Oh, Hee-Kyun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.48 no.5
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    • pp.303-308
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    • 2022
  • Objectives: Selection of treatment methods for mandibular condylar fractures remains controversial. In this study, we investigated treatment methods for condylar fractures to determine the indications for open or closed reduction. Patients and Methods: Patients >12 years of age treated for mandibular condylar fractures with a follow-up period of ≥3 months were included in this study. The medical records of enrolled patients were reviewed for sex, age, fracture site, treatment method (open or closed reconstruction), postoperative intermaxillary fixation period, operation time, and complications. Radiological analysis of fracture fragment displacement and changes in ramal height difference was performed using computed tomography and panoramic radiography. Results: A total of 198 patients was investigated, 48.0% (n=95) of whom underwent closed reduction and 52.0% (n=103) underwent open reduction. There was no significant correlation between reduction method and patient sex, age, or follow-up period. No statistically significant difference between the incidence of complications and treatment method was observed. None of the patients underwent open reduction of condylar head fracture. Binary logistic regression analysis showed that open reduction was significantly more frequent in patients with subcondylar fracture compared to in those with a fracture in the condylar head area. There was no statistically significant correlation between the groups and fracture fragment displacement. However, there was a significant difference between the treatment groups in amount of change in ramal height difference between the fractured and the non-fractured sides during treatment. Conclusion: No significant clinical differences were found between the open and closed reduction methods in patients with mandibular condylar fractures. According to fracture site, closed reduction was preferred for condyle head fractures. There was no significant relationship between fracture fragment displacement and treatment method.

Tissue Distribution, SNP Detection and Association Study with Immune Traits of Porcine LBP and CD14 Genes

  • Liu, H.Z.;Li, X.Y.;Liu, B.;Yu, M.;Ma, Y.H.;Chu, M.X.;Li, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.8
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    • pp.1080-1087
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    • 2008
  • Lipopolysaccharide binding protein (LBP) and CD14 protein play important roles in the defense against infection of Gram-negative bacteria. In the present study, tissue distribution and polymorphism of porcine LBP and CD14 genes were analyzed. Real-time PCR results showed that the porcine LBP gene was especially highly expressed in liver, while CD14 gene was highly expressed in liver and spleen tissues. A 1,732 bp cDNA fragment of porcine LBP gene and a 1,682 bp genomic DNA fragment of CD14 gene were isolated. Polymorphisms were identified in these two fragments and showed that there were 14 potential SNPs in the porcine LBP gene and 3 potential SNPs in the porcine CD14 gene. Three SNPs, 292G/A (Gly/Ser), 1168G/A (Ala/Thr) of the LBP gene and -61G/A of the CD14 gene, were genotyped using restriction fragment length polymorphism (RFLP) method. Association analyses indicated that polymorphism of the 292G/A locus was significantly associated with porcine immune traits hematocrit (HCT), IgG and delayed-type hypersensitivity (DTH) (p<0.01), and the 1168G/A locus was significantly associated with HCT and mean corpuscular volume (MCV) traits (p<0.05). No significant association was found between the -61G/A locus and immune traits of the pig. Our data indicated that the LBP gene was significantly associated with immune traits of pig. Also, we identified some SNPs which may be useful markers for disease-resistant breeding of pigs.

Characterization of PCR fragment of metallothionein gene from liver mRNS of channel catfish (챠넬메기의 간 mRNA 로부터 분리한 metallothionein 유전자의 PCR 절편의 특성)

  • Song, Young-Hwan
    • Journal of fish pathology
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    • v.10 no.1
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    • pp.39-44
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    • 1997
  • Metallothionein is an important and essential protein to control the intracellular concentration of heavy metals, which exist in all organisms from bacteria to vertebrates. Although the detailed functions and induction mechanisms of metallothionein gene have not been clearly characterized until yet, the structure of several metallothionein genes has been revealed. Especially, piscine metallothionein is regarded as an important protein because it is induced by several heavy metal pollutants and environmental stress and it could be determined the comparative amount of heavy metals and the extent of environmental stress by assaying the RNA transcript of metallothionein gene in the method of the quantitative RT-PCR(Reverse Transcriptase Polymerase Chain Reaction). In this study I have characterized the 450 bp PCR fragment of metallothionein gene amplified by using the mixture of internal specific primers and universal 3' end primer. The nucleotide sequence analysis of 450 bp PCR fragment amplified in cDNA library of channel catfish did not show strong homology to other piscine metallothionein genes.

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Anatomical Locking Plate with Additional K-wire Fixation for Distal Clavicle Fracture

  • Nam, Woo-Dong;Moon, Sung-Hoon;Choi, Ki-Yong
    • Clinics in Shoulder and Elbow
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    • v.20 no.4
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    • pp.230-235
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    • 2017
  • Background: Neer type II distal clavicle fractures have the drawback of coracoclavicular instability and insufficient distal bony fragment, thereby making it difficult to achieve adequate fixation. Although various surgical treatments have been described for Neer type II fracture, the optimal treatment remains controversial. This study reports the clinical results and usefulness of anatomical locking plate with additional K-wire fixation. Methods: A totally of 21 patients with type II distal clavicle fracture were included in the study. The surgical procedure reduced the fracture temporarily; it included insertion of one or two K-wire from the lateral margin of the distal fragment to the proximal fragment through the fracture site, followed by application and fixation of the locking plate. The bony union and migration of K-wire was evaluated in the follow-up radiography. The coracoclavicular distance and acromioclavicular joint arthrosis were assessed at the final follow-up. The Constant Score (CS) and Korean Shoulder Score (KSS) were evaluated for clinical scoring. Results: Bone union was achieved in all cases. At the final follow-up, coracoclavicular distance of the injured shoulder was increased, as compared to the intact shoulder (p=0.002), with no accompanying clinical symptoms. No K-wire migration was observed. At the final follow-up, K-wire irritation was observed in two cases and acromioclavicular arthrosis in one case, with no other adverse effects. Pain visual analogue scale, CS, and KSS were improved in all cases. Conclusions: The method of anatomical locking plate with additional K-wire fixation could be useful in achieving beneficial clinical results.

Genetic Diversity of Didymella bryoniae for RAPD Profiles Substantiated by SCAR Marker in Korea

  • Shim, Chang-Ki;Seo, Il-Kyo;Jee, Hyeong-Jin;Kim, Hee-Kyu
    • The Plant Pathology Journal
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    • v.22 no.1
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    • pp.36-45
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    • 2006
  • Twenty isolates of Didymella bryoniae were isolated from infected cucurbit plants in various growing areas of southern Korea in 2001 and 2002. Random Amplified Polymorphic DNA (RAPD) group [RG] I of D. bryoniae was more virulent than RG IV to watermelon. Virulence of the RG I isolate was strong to moderate to cucumber, whereas that of the RG IV varied from strong, moderate to weak. Two hundred seventy-three amplified fragments were produced with 40 primers, and were analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYSPC. At the distance level of 0.7, two major genomic DNA RAPD groups were differentiated among 20 isolates. The RG I included 7 isolates from watermelon and one isolate from melon, whereas the RG IV included 12 isolates from squash, cucumber, watermelon and melon. Amplification of internal transcribed spacer (ITS) region and small subunit rRNA region from the 20 isolates yielded respectively a single fragment. Restriction pattern with 12 restriction enzymes was identical for all isolates tested, suggesting that variation in the ITS and small subunit within the D. bryoniae were low. Amplification of the genomic DNAs of the tested isolates with the sequence characterized amplified regions (SCAR) primer RG IF-RG IR specific for RG I group resulted in a single band of 650bp fragment for 8 isolates out of the 20 isolates. Therefore, these 8 isolates could be assigned into RG I. The same experiments done with RG IIF-RG IIR resulted in no amplified PCR product for the 20 isolates tested. An about 1.4 kb-fragment amplified from the RG IV isolates was specifically hybridized with PCR fragments amplified from genomic DNAs of the RG IV isolates only, suggesting that this PCR product could be used for discriminating the RG IV isolates from the RG I isolates as well other fungal species.

Identification of Medicinal Mushroom Species Based on Nuclear Large Subunit rDNA Sequences

  • Lee Ji Seon;Lim Mi Ok;Cho Kyoung Yeh;Cho Jung Hee;Chang Seung Yeup;Nam Doo Hyun
    • Journal of Microbiology
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    • v.44 no.1
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    • pp.29-34
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    • 2006
  • The purpose of this study was to develop molecular identification method for medical mushrooms and their preparations based on the nucleotide sequences of nuclear large subunit (LSD) rDNA. Four specimens were collected of each of the three representative medicinal mushrooms used in Korea: Ganoderma Incidum, Coriolus versicolor, and Fomes fomentarius. Fungal material used in these experiments included two different mycelial cultures and two different fruiting bodies from wild or cultivated mushrooms. The genomic DNA of mushrooms were extracted and 3 nuclear LSU rDNA fragments were amplified: set 1 for the 1.1-kb DNA fragment in the upstream region, set 2 for the 1.2-kb fragment in the middle, and set 3 for the 1.3-kb fragment downstream. The amplified gene products of nuclear large subunit rDNA from 3 different mushrooms were cloned into E. coli vector and subjected to nucleotide sequence determination. The sequence thus determined revealed that the gene sequences of the same medicinal mushroom species were more than $99.48\%$ homologous, and the consensus sequences of 3 different medicinal mushrooms were more than $97.80\%$ homologous. Restriction analysis revealed no useful restriction sites for 6-bp recognition enzymes for distinguishing the 3 sequences from one another, but some distinctive restriction patterns were recognized by the 4-bp recognition enzymes AccII and HhaI. This analysis was also confirmed by PCR-RFLP experiments on medicinal mushrooms.

Bacterial ${\beta}$-Lactamase Fragment Complementation Strategy Can Be Used as a Method for Identifying Interacting Protein Pairs

  • Park, Jong-Hwa;Back, Jung-Ho;Hahm, Soo-Hyun;Shim, Hye-Young;Park, Min-Ju;Ko, Sung-Il;Han, Ye-Sun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.10
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    • pp.1607-1615
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    • 2007
  • We investigated the applicability of the TEM-l ${\beta}$-lactamase fragment complementation (BFC) system to develop a strategy for the screening of protein-protein interactions in bacteria. A BFC system containing a human Fas-associated death domain (hFADD) and human Fas death domain (hFasDD) was generated. The hFADD-hFasDD interaction was verified by cell survivability in ampicillin-containing medium and the colorimetric change of nitrocefin. It was also confirmed by His pull-down assay using cell lysates obtained in selection steps. A coiled-coil helix coiled-coil domain-containing protein 5 (CHCH5) was identified as an interacting protein of human uracil DNA glycosylase (hUNG) from the bacterial BFC cDNA library strategy. The interaction between hUNG and CHCH5 was further confirmed with immunoprecipitation using a mammalian expression system. CHCH5 enhanced the DNA glycosylase activity of hUNG to remove uracil from DNA duplexes containing a U/G mismatch pair. These results suggest that the bacterial BFC cDNA library strategy can be effectively used to identify interacting protein pairs.