• Title/Summary/Keyword: 6S rDNA analysis

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The Diversity of Heterotrophic Bacteria Isolated from Intestine of Starfish(Asterias amurensis) by Analysis of 16S rDNA Sequence (16S rDNA염기서열에 의한 불가사리(Asterias amurensis) 장내에서 분리된 종속영양세균 군집의 다양성)

  • Choi, Gang-Guk;Lee, Oh-Hyung;Lee, Geon-Hyoung
    • The Korean Journal of Ecology
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    • v.26 no.6
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    • pp.307-312
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    • 2003
  • To study the diversity of heterotrophic bacteria isolated from intestine of starfish, Asterias amurensis, we collected starfishes from the coastal area near Jangheung-Gun, Jeollanam-Do, Korea during July, 2000. Population density and bacterial diversity in the intestine of starfish were measured. The results were as follows; The population densities of heterotrophic bacteria in the intestine of starfish were 8.65${\pm}$0.65${\times}10^3\;dfu\;g^{-1}$. Gram positive bacteria occupied 59% among 29 isolates. The community structure of dominant heterotrophic bacteria in the intestine of starfish consisted of Bacillaceae in the low G+C gram positive bacteria subphylum, Microbacteriaceae in the high G+C gram positive bacteria subphylum, and Alteromonadaceae in ${\gamma}$-Proteobacteria subphylum. Among eight strains of Bacillus spp., three strains showed more than 97% identity, but five strains showed about 90% identity with type strain on the basis of partial 16S rDNA sequence.

Estimation of Distribution of a Commensal Thermophile in Soil by Competitive Quantitative PCR and Terminal Restriction Fragment Length Polymorphism Analysis

  • Rhee, Sung-Keun;Hong, Seung-Pyo;Bae, Jin-Woo;Jeon, Che-Ok;Lee, Seung-Goo;Song, Jae-Jun;Poo, Ha-Ryoung;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.940-945
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    • 2001
  • Symbiobacterium toebii has been previously reported as a novel commensal thermophile exhibiting a commensal interaction with thermophilic Geobacillus sp. SK-1. We investigated the distribution of this commensal thermophile in various soils using molecular methods, such as quantitative PCR and terminal restriction fragment polymorphism analysis. Based on a nested competitive quantitative PCR the 16S rDNA of the commensal thermophile was only detected in compost soils at about $1.0{\times}10^4$ cpoies per gram of soil, corresponding to $0.25{\times}10^4$ cells per gram of soil. However, in an enrichment experiment at $60^{\circ}C$, about $1.0{\times}10^8$ copies of 16S rDNA molecules were detected per ml of enriched culture broth for all the soils, and more than 0.1 mM indole accumulated as the product of commensal bacterial growth. When incubated at $30^{\circ}C$, neither the 16S rDNA of the commensal bacterium nor any indole accumulation was detected. Accordingly, even though the 16S rDNA of the bacterium was only detected in the compost soils by a nested PCR, the presence of the 16S rDNA molecules of commensal thermophile and accumulation of indole in all the enriched cultures appeared to indicate that the commensal thermophile is widely distributed in various soils.

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16S rDNA Analysis 9f Bacterial Diversity in Three Fractions of Cow Rumen

  • Cho, Soo-Jeong;Cho, Kye-Man;Shin, Eun-Chule;Lim, Woo-Jin;Hong, Su-Young;Choi, Byoung-Rock;Kang, Jung-Mi;Lee, Sun-Mi;Kim, Yong-Hee;Kim, Hoon;Yun, Han-Dae
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.92-101
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    • 2006
  • The bacterial diversity of the bovine rumen was examined using a PCR-based approach. 16S rDNA sequences were amplified and cloned from three fractions of rumen (solid, fluid, and epithelium) that are likely to represent different bacterial niches. A total of 113 clones were sequenced, and similarities to known l6S rDNA sequences were examined. About $47.8\%$ of the sequences had $90-97\%$ similarity to 16S rDNA database sequences. Furthermore, about $62.2\%$ of the sequences were $98-100\%$ similar to 16S rDNA database sequences. For the remaining $6.1\%$, the similarity was less than $90\%$. Phylogenetic analysis was also used to infer the makeup of the bacterial communities in the different rumen fractions. The Cytophaga-Flexibacter-Bacteroides group (CFB, $67.5\%$), low G+C Gram-positive bacteria (LGCGPB, $30\%$), and Proteobacteria $(2.5\%)$ were represented in the rumen fluid clone set; LGCGPB $(75.7\%)$, CFB$(10.8\%)$, Proteobacteria $(5.4\%)$, high G+C Gram-positive bacteria (HGCGPB, $5.4\%$), and Spirochaetes $(2.7\%)$ were represented in the rumen solid clone set; and the CFB group $(94.4\%)$ and LGCGPB $(5.6\%)$ were represented in the rumen epithelium clone set. These findings suggest that the rumen fluid, solid, and epithelium support different microbial populations that may play specific roles in rumen function.

A Method for Comparing Multiple Bacterial Community Structures from 16S rDNA Clone Library Sequences

  • Hur, Inae;Chun, Jongsik
    • Journal of Microbiology
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    • v.42 no.1
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    • pp.9-13
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    • 2004
  • Culture-independent approaches, based on 16S rDNA sequences, are extensively used in modern microbial ecology. Sequencing of the clone library generated from environmental DNA has advantages over fingerprint-based methods, such as denaturing gradient gel electrophoresis, as it provides precise identification and quantification of the phylotypes present in samples. However, to date, no method exists for comparing multiple bacterial community structures using clone library sequences. In this study, an automated method to achieve this has been developed, by applying pair wise alignment, hierarchical clustering and principle component analysis. The method has been demonstrated to be successful in comparing samples from various environments. The program, named CommCluster, was written in JAVA, and is now freely available, at http://chunlab.snu.ac.kr/commcluster/.

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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    • v.15 no.2
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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.

Identification of Fusobacterium nucleatum isolated from Korean by F. nucleatum subspecies-specific DNA probes (Dot blot hybridization법을 이용한 Fusobacterium nucleatum 아종-특이 DNA 프로브의 특이성 평가)

  • Kim, Hwa-Sook;Kook, Joong-Ki
    • Journal of Korean society of Dental Hygiene
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    • v.6 no.4
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    • pp.311-324
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    • 2006
  • The purpose of this investigation was to evaluate of the specificity of Fusobacterium nucleatum subspecies-specific DNA probes using dot blot hybridization. To confirm whether the clinical isolates were F. nucleatum or not, 16S rDNA of them were cloned and sequenced. The sequencing data were used in homology search with database of GenBank. When the homology was above 98% compared with the nucleotide sequence of a certain bacteria, it was judged as the same species with the bacteria. 23 strains of F. nucleatum were isolates from subgingival plaque of periodontitis patient. The clinical isolates of F. nucleatum were classified into 10 groups using phylogenetic analysis of 16S rDNA sequence. F. nucleatum subspecies nucleatum-specific DNA probe Fu4(1.3 kb) reacted with genomic DNAs from 8 type strains of F. nucleatum and it reacted strongly with those from 8 clinical isolates. The Fp4(0.8 kb) reacted with F. nucleatum subsp. polymorphum ATCC 10953 and one clinical isolates. Fv35(1.9 kb) and Fs17(8.2 kb) probes reacted with genomic DNAs from F. nucleatum subsp. vincentii ATCC 49256 and F. nucleatum subsp. fusiform ATCC 51190, respectively. Our results showed that it is not enough to evaluate the specificity of F. nucleatum subspecies-specific DNA probes with only dot blot hybridization. Therefore, Southern blot analysis will be necessary to confirm the specificity of F. nucleatum subspecies-specific DNA probes.

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Emendation of Rhodomonas marina (Cryptophyceae): insights from morphology, molecular phylogeny and water-soluble pigment in an Arctic isolate

  • Niels Daugbjerg;Cecilie B. Devantier
    • ALGAE
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    • v.39 no.2
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    • pp.75-96
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    • 2024
  • Rhodomonas (Cryptophyceae) and species assigned to this genus have undergone numerous taxonomic revisions. This also applies to R. marina studied here as it was originally assigned as a species of Cryptomonas and later considered a variation of R. baltica, the type species. Despite being described more than 130 years ago, R. marina still lacks a comprehensive characterization. Light and electron microscopy were employed to delineate a strain from western Greenland. The living cells were 18 ㎛ long and 9 ㎛ wide, elliptical in shape with a pointed to rounded posterior and truncated anterior in lateral view. Two sub-equal flagella emerged from a vestibulum, where also a furrow extended. In transmission electron microscopy, the furrow was associated with a tubular gullet and the pyrenoid embedded in a deeply lobed chloroplast. The chloroplast contained DNA in perforations and was surrounded by starch grains. A tubular nucleomorph was enclosed within the pyrenoid matrix. In scanning electron microscopy, the inner periplast consisted of rectangular plates with rounded edges and posteriorly these were replaced by a sheet-like structure. The water-soluble pigment was Crypto-Phycoerythrin type I (Cr-PE 545). A phylogenetic inference based on SSU rDNA confirmed the identity of strain S18 as a species of Rhodomonas as it clustered with congeners but also Rhinomonas, Storeatula, and Pyrenomonas. These genera formed a monophyletic clade separated from a diverse assemblage of other cryptophyte genera. To further explore the phylogeny of R. marina a concatenated phylogenetic analysis based on the SSU rDNA-ITS1-5.8S rDNA-ITS2-LSU rDNA region was performed but included only closely related species. The secondary structure of nuclear internal transcribed spacer 2 was predicted and compared to similar structures in related species. Using morphological and molecular signatures as diagnostic features the description of R. marina was emended.

Heterogeneity Analysis of the 16S rRNA Gene Sequences of the Genus Vibrio (Vibrio 속 16S rRNA 유전자 염기서열의 이질성 분석)

  • Ki, Jang-Seu
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.430-434
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    • 2009
  • Bacterial 16S rRNA gene sequences have been widely used for the studies on molecular phylogeny, evolutional history, and molecular detections. Bacterial genomes have multiple rRNA operons, of which gene sequences sometimes are variable. In the present study, heterogeneity of the Vibrio 16S rRNA gene sequences were investigated. Vibrio 16S rRNA sequences were obtained from GenBank databases, considering the completion of gene annotation of Vibrio genome sequences. These included V. cholerae, V. harveyi, V. parahaemolyticus, V. splendidus, and V. vulnificus. Chromosome 1 of the studied Vibrio had 7~10 copies of the 16S rRNA gene, and their intragenomic variations were less than 0.9% dissimilarity (more than 99.1% DNA similarity). Chromosome 2 had none or single 16S rRNA gene. Intragenomic 16S rRNA genotypes were detected at least 5 types (V. vulnificus #CMCP6) to 8 types (V. parahaemolyticus #RIMD 2210633, V. harveyi #ATCC BAA-1116). These suggest that Vibrio has high heterogeneity of the 16S rRNA gene sequences.

Cloning of the Bombyx mori short neuropeptide F receptor (BsNPF-R) cDNA (누에 short neuropeptide F receptor (BsNPF-R)의 cDNA cloning)

  • Shin, Hyojung;Kwon, Kisang;Hong, Sun Mee;Kim, Hong Geun;Park, Kwan-Ho;Choi, Ji-Young;Kim, Seung-Whan;Yu, Kweon;Kwon, O-Yu
    • Journal of Life Science
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    • v.26 no.6
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    • pp.721-726
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    • 2016
  • It has already been reported that short neuropeptide F (sNPF) stimulates feeding behaviors in a wide variety of insect species. In the present study, we cloned cDNA, encoding a sNPF receptor homologue from a silkworm, Bombyx mori, named BsNPF-R. The amino acid sequence of BsNPF-R was compared with those of sNPF-R thus far reported, which is shared with humans (36%), mice (34%), zebrafish (35%), and fruit flies (51%), respectively. A BsNPF-R protein’s mass was theoretically estimated to be 42,731 Da and it is a putative plasma membrane-penetrating protein. The mRNA expression of BsNPF-R was tested; the results showed that a strong expression was detected at the midgut, post-silk gland, Malpighian, and testis; however, a weak expression was at the fat body, hemocyte, and ovary. In addition, the synthesized sNPF of a silkworm regulated the BsNPF-R mRNA expression through the cell-based functional analysis.

mtDNA Diversity and Phylogenetic Analysis of Korean Native Goats (한국재래염소의 mtDNA 다양성 및 계통유전학적 분석)

  • Kim, Jae-Hwan;Cho, Chang-Yeon;Choi, Seong-Bok;Cho, Young-Moo;Yeon, Seung-Hum;Yang, Boh-Suk
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1329-1335
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    • 2011
  • Korean native goats, which are characterized by black coat color, have existed on the Korean peninsula for a long time. Until now, there has been no comprehensive investigation concerning their genetic diversity, phylogenetic analysis or origin. In this study, we investigated the genetic diversity and verified phylogenetic status of the Korean native goat using the 453-bp fragment of the hypervariable fragment I (HVI) of mitochondrial DNA (mtDNA) D-loop region from 60 individuals among 5 populations. The Korean native goat showed less haplotype diversity when compared with goats from other countries. In addition, 6 haplotypes that had not been previously reported were verified in this study. In phylogenetic analyses with other country's goats, 10 haplotypes from Korean native goats were classified into mtDNA lineage A. Moreover, in a phylogenetic tree for goats which contained mtDNA lineage A, 8 of 10 haplotypes could be included in a subgroup with goats from Vietnam and an area of China. However, none of the remaining haplotypes belonged to a major group of Korean native goats and were located on different independent positions. These results suggest that almost Korean native goats aligned more closely to China and Vietnam breeds in mtDNA lineage A and there was no gene flow from other mtDNA lineages. Our results will contribute to conservation strategies and genetic breeding of Korean native goats.