• 제목/요약/키워드: DNA Sequencing

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정수처리에서 생물학적 망간처리 (Biological Manganese Removal in Water Treatment)

  • 김범수;윤재경;안효원;김충환
    • 상하수도학회지
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    • 제20권1호
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    • pp.44-52
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    • 2006
  • Bio-filtration processes using honeycomb tubes (process 1) and aeration and manganese-sand filtration (process 2) were evaluated for the biological manganese removal efficiency. The concentration of manganese at effluent was stabilized after 20days operation in process 1. It was estimated the required time for attaching and growing microorganisms to honeycomb tubes. In long term of operation periods, manganese removal efficiency was dropped for the excessively attached biofilm and manganese dioxide to honeycomb tubes. It took several days for normal operation in process 2, after that manganese removal efficiency was increased to 98% and stabilized for 1.5 years. Microorganisms in process 1 and 2 were isolated and cultured to characterize manganese-oxidizing bacteria. Among the four types of colony, light brown colony was turned blue color by leuco crystal violet spot test. Stenotropomonas genus, known as manganese-oxidizing bacteria, was identified by 16S rDNA partial sequencing analysis which was isolated in process 1 and 2. For the biological treatment to remove manganese, these two considerations are important. One is to choose the proper media attaching manganese oxidant, another one is to define the cultural condition of isolated manganese-oxidizing bacteria.

Refinements for the amplification and sequencing of red algal DNA barcode and RedToL phylogenetic markers: a summary of current primers, profiles and strategies

  • Saunders, Gary W.;Moore, Tanya E.
    • ALGAE
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    • 제28권1호
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    • pp.31-43
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    • 2013
  • This review provides a comprehensive summary of the PCR primers and profiles currently in use in our laboratory for red algal DNA barcoding and phylogenetic research. While work focuses on florideophyte taxa, many of the markers have been applied successfully to the Bangiales, as well as other lineages previously assigned to the Bangiophyceae sensu lato. All of the primers currently in use with their respective amplification profiles and strategies are provided, which can include full fragment, overlapping fragments and what might best be called "informed overlapping fragments", i.e., a fragment for a marker is amplified and sequenced for a taxon and those sequence data are then used to identify the best primers to amplify the remaining fragment(s) for that marker. We extend this strategy for the more variable markers with sequence from the external PCR primers used to "inform" the selection of internal sequencing primers. This summary will hopefully serve as a useful resource to systematists in the red algal community.

One Step Cloning of Defined DNA Fragments from Large Genomic Clones

  • Scholz, Christian;Doderlein, Gabriele;Simon, Horst H.
    • BMB Reports
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    • 제39권4호
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    • pp.464-467
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    • 2006
  • Recently, the nucleotide sequences of entire genomes became available. This information combined with older sequencing data discloses the exact chromosomal location of millions of nucleotide markers stored in the databases at NCBI, EMBO or DDBJ. Despite having resolved the intron/exon structures of all described genes within these genomes with a stroke of a pen, the sequencing data opens up other interesting possibilities. For example, the genomic mapping of the end sequences of the human, murine and rat BAC libraries generated at The Institute for Genomic Research (TIGR), reveals now the entire encompassed sequence of the inserts for more than a million of these clones. Since these clones are individually stored, they are now an invaluable source for experiments which depend on genomic DNA. Isolation of smaller fragments from such clones with standard methods is a time consuming process. We describe here a reliable one-step cloning technique to obtain a DNA fragment with a defined size and sequence from larger genomic clones in less than 48 hours using a standard vector with a multiple cloning site, and common restriction enzymes and equipment. The only prerequisites are the sequences of ends of the insert and of the underlying genome.

Ribosomal DNA의 ITS부위에 대한 RFLP 분석에 의한 Phellinus baumii PMO-P4의 유전학적 특성 (Genetic characterization of Phellinus baumii PMO-P4 by analyzing restriction fragment length polymorphisms of nuclear ribosomal DNA internal transcribed spacers (ITS))

  • 장윤희;김태락;김현수;여익현;이상윤;하효철
    • 한국버섯학회지
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    • 제4권2호
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    • pp.43-47
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    • 2006
  • 국내에서 재배하여 생산되고 있는 상황버섯의 일종인 PMO-P4균주에 대한 ITS 영역의 염기서열 분석을 실시하였으며 목질 진흙버섯으로 잘 알려져 있는 P. linteus와 함께 RFLP분석을 통하여 상호 비교한 결과 PMO-P4균주는 P. baumii로 판명되었다. 이 결과를 토대로 이미 보고되어 있는 Phellinus속 균주들과의 종간 ITS 영역의 상동성을 비교한 결과 48.6%-72.2%였으며 본 연구에서 비교한 종들 가운데서는 P. linteus와 상동성이 가장 높았으며 P. gilvus와 상동성이 가장 낮았다.

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Distribution of Dominant Bifidobacteria in the Intestinal Microflora of Korean Adults and Seniors, Identified by SDS-PAGE of Whole Cell Proteins and 16S rDNA Sequence Analysis

  • KIM TAE WOON;SONG HEE SUNG;KIM HAE YEONG
    • Journal of Microbiology and Biotechnology
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    • 제15권2호
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    • pp.388-394
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    • 2005
  • In order to investigate the distribution of dominant Bifidobacterium species in intestinal microflora of Korean adults and seniors, SDS-PAGE profiles of whole cell proteins were used for the identification of bifidobacteria. To confirm the reliability of SDS-PAGE, the Bifidobacterium species identified by SDS-PAGE of whole cell proteins were validated by using 16S rDNA sequencing analysis. The results of SDS­PAGE corresponded well with those determined by the analysis of 16S rDNA sequencing. Based on the analysis of SDS-PAGE patterns on unidentified fecal strains which showed positive in fructose-6-phosphate phosphoketolase activity, B. adolescentis, B. longum, and B. bifidum were identified in the feces of adults, and B. adolescentis, B. longum, B. bifidum, B. breve, and B. dentium were identified in those of seniors. In most of the fecal samples tested, the predominant Bifidobacterium species consisted of only a few species, and differences in the distribution and numbers of Bifidobacterium species were observed between adults and seniors. B. adolescentis and B. longum were found to be the most common species in feces of adults, but not in seniors. Accordingly, the distribution and abundance of bifidobacteria in the human intestinal microflora varied depending on the age of hosts.

활성슬러지내의 전기화학적활성 박테리아 분포 특성 (Distribution of Electrochemically Active Bacteria in Activated Sludge Characteristics)

  • 손형식;손희종;김미아;이상준
    • KSBB Journal
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    • 제26권5호
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    • pp.407-411
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    • 2011
  • Microbial fuel cell (MFC) wes enriched using sludge in wastewater treatment. The microbial community of activated sludge and enriched MFC were analyzed by FISH (fluorescent in situ hybridization) and 16S rDNA sequencing. Bacteroidetes group were pre-dominant in activated sludge by FISH. ${\alpha}$ group, ${\gamma}$ group and Acintobacter group were dominant and they were similar to distribution. The average value of 10 peak of MFC is 0.44C. When MFC wase enriched by sludge, ${\gamma}$-Proteobacteria, Plantomycetes group increased 70% and 60%, respectively. In results of 16S rDNA sequencing, Sphiringomonas sp. was comprised in ${\alpha}$ proteobacteria and Enterobacter sp., Klebsiella sp., Acinetobacter sp., Bacillus sp. were comprised in ${\gamma}$ proteobacteria and Chryseobacterium sp. was comprised in Flavobacteria were isolated from sludge.

소아의 치아 우식 부위별 세균 다양성 (Bacterial diversity in children's dental caries)

  • 김은미;백근식;하명옥
    • 한국치위생학회지
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    • 제13권5호
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    • pp.889-900
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    • 2013
  • Objectives : Molecular biology techniques were employed to assess diversity of bacterial in children's dental caries. Methods : DNA of germs was extracted and the diversity of the 16S rRNA clones was analyzed by amplified rDNA restriction analysis and sequencing. The experimental samples were pit and fissure caries (PC), deep dentinal caries (DC), smooth surface caries (SC), and supragingival plaque (PQ) from 50 children of age less than 12 years old. The control group was healthy teeth supragingival plaque (HT). Thirty clones from each 16S rRNA clone library of 5 samples were randomly selected, thus a total of 150 clones were analyzed. Results : Amplified rDNA restriction analysis uncovered 18, 20, 11, 17, and 22 phylotypes from healthy teeth, pit and fissure caries, deep dentinal caries, smooth surface caries, and supragingival plaque, respectively. Sequencing analysis found the dominance of Actinomycs naeslundii and Fusobacterium nucleatum in the healthy teeth; Leptotrichia sp. in the pit and fissure caries; Actinomyces sp., Streptococcus mutans, and Rahnella aquatilis in the deep dentinal caries; Streptococcus mutans and Actinomyces sp. in the smooth surface caries; Enterobacter hormaechei and Streptococcus sanguinis in the supragingival plaque. Conclusions : Clonal analysis identified 6 phyla, 20 genera, and 51 species.

DNA Sequencing and Expression of the Circumsporozoite Protein of Plasmodium vivax Korean Isolate in Escherichia coli

  • Lee, Hyeong-Woo;Lee, Jong-Soo;Lee, Won-Ja;Lee, Ho-Sa
    • Journal of Microbiology
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    • 제37권4호
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    • pp.234-242
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    • 1999
  • To obtain the recombinant circumsporozoite (CS) protein for the diagnosis of patients and seroepidemiology of Plasmodium vivax malaria which have been prevalent in northern part of Kyonggido, the CS protein gene was amplified by the polymerase chain reaction (PCR) from genomic DNA of the Korean vivax malaria patient. The gene consists of 1,123 nucleotides except signal peptide sequences and had an uninterrupted reading frame encoding a protein of 374 amino acids with a central region of 20 tandem repeats of the nonapeptide. The CS protein gene was expressed in Escherichia coli and purified, the molecular weight of recombinant CS protein was about 44 kDa (monomer) under denaturing purification and about 65 kDa (dimer) under native purification by SDS-PAGE. The purified recombinant CS protein which has antigenicity to malaria patients in Western blot analysis and Enzyme-linked immunosorbent assay, reacted only with the serum of P. vivax (PV210) infected malaria patients with no cross reaction to the P. falciparum malaria patient. The recombinant CS protein purified in this study will serve as a useful antigen to support the diagnosis of malaria patients and seroepidemiology.

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병원성장내세균에서 phoP-phoQ operon의 지배를 받는 phoA 유전자의 cloning 및 염기서열결정 (Cloning and Sequencing of the phoA Gene which is Regulated by the phoP-phoQ Operon in Pathogenic Enteric Bacteria)

  • 김성광;이태윤
    • Journal of Yeungnam Medical Science
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    • 제12권2호
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    • pp.237-245
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    • 1995
  • Klebsiella pneumoniae 의 phoA 유전자를 함유하는 DNA를 plasmid pACYC184에 클로닝 하였다. 클로닝된 DNA의 크기는 4.0 kb이었으며 제한효소지도를 작성한 결과 3개의 PstI 절단부위와 4개의 PvuII 절단부위가 발견되었다. Klebsiella pneumoniae 의 phoA 유전자의 염기서열은 Escherichia coli와 매우 유사하여 80%의 유사성을 보였으며 이는 이 두 균종이 서로 유전적으로 매우 가까운 관계에 있음을 시사하였다.

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The First Acanthamoeba keratitis Case of Non-Contact Lens Wearer with HIV Infection in Thailand

  • Tananuvat, Napaporn;Techajongjintana, Natnaree;Somboon, Pradya;Wannasan, Anchalee
    • Parasites, Hosts and Diseases
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    • 제57권5호
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    • pp.505-511
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    • 2019
  • Acanthamoeba keratitis (AK) is a rare sight-threatening corneal infection, often reporting from contact lens wearers. An asymptomatic human immunodeficiency virus (HIV)-infected Thai male without history of contact lens use complained foreign body sensation at his left eye during motorbike riding. He had neither specific keratitis symptoms nor common drugs responding, which contributed to delayed diagnosis. By corneal re-scraping, Acanthamoeba-like cysts were detected by calcofluor white staining and agar culture. The etiological agent obtained from the culture was molecularly confirmed by Acanthamoeba spp.-specific PCR, followed by DNA sequencing. The results from BLAST and phylogenetic analysis based on the DNA sequences, revealed that the pathogen was Acanthamoeba T4, the major genotype most frequently reported from clinical isolates. The infection was successfully treated with polyhexamethylene biguanide resulting in corneal scar. This appears the first reported AK case from a non-contact lens wearer with HIV infection in Thailand. Although AK is sporadic in developing countries, a role of free-living Acanthamoeba as an opportunistic pathogen should not be neglected. The report would increase awareness of AK, especially in the case presenting unspecific keratitis symptoms without clinical response to empirical antimicrobial therapy.