• Title/Summary/Keyword: rRNA sequence

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Cloning and Sequence Analysis of the Aminoglycoside Resistance Gene from a Nebramycin Complex Producer, Streptoalloteichus hindustanus

  • Hyun, Chang-Gu;Kim, Jong-Woo;Han, Jae-Jin;Choi, Young-Nae;Suh, Joo-Won
    • Journal of Microbiology and Biotechnology
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    • v.8 no.2
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    • pp.146-151
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    • 1998
  • The aminoglycoside multiple-resistance determinant from Streptoalloteichus hindustanus was cloned into Streptomyces lividans and named nbrB. The 1.2-kb ApaI- BclI fragment encompassing nbrB was located within a 2.6-kb ApaI fragment by successive subcloning experiments. The complete DNA nucleotide sequence of 1.2-kb containing nbrB was determined. The sequence contains an open reading frame that putatively encodes a polypeptide of 281 amino acids with a predicted molecular weight of 30,992. The deduced amino acid sequence of nbrB shows identities of 85.1% to kgmB of S. tenebrarius, 59.6% to sgm of Micromonospora zionensis, and 57.7% to grm of M. rosea. The similarity of nbrB to kgmB suggests that nbrB encodes a 16S rRNA methylase similar to that encoded by kgmB and that both genes might be derived from a common ancestral gene.

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Cloning and Expression of A Bacillus licheniformis Cellulase Gene (Bacillus licheniformis WL-12의 cellulase 유전자 클로닝과 발현)

  • Yoon, Ki-Hong
    • Korean Journal of Microbiology
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    • v.42 no.4
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    • pp.313-318
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    • 2006
  • A thermophilic bacterium producing the extracellular cellulase was isolated from soybean paste, and the isolate WL-12 has been identified as Bacillus licheniformis on the basis on its 16S rRNA sequence, morphology and biochemical properties. A gene encoding the cellulase of B. licheniformis WL-12 was cloned and its nucleotide sequence was determined. This cellulase gene, designated celA, consisted of 1,551 nucleotides, encoding a polypeptide of 517 amino acid residues. The gene product contained catalytic domain and cellulose binding domain. The deduced amino acid sequence was highly homologous to those of cellulases of B. licheniformis, B. subtilis and B. amytoliquefaciens belonging to the glycosyl hydrolase family 5. When the celA gene was highly expressed using a strong B. subtilis promoter, the extracellular cellulase was produced up to 7.0 units/ml in B. subtilis WB700.

Microbial Community Analysis Isolated from Red Starfish (Certonardoa semiregularis) Gut (빨강불가사리(Certonardoa semiregularis)에서 분리된 세균의 군집구조 분석)

  • Lee, Hae-Ri;Park, So-Hyun;Kim, Dong-Hwi;Moon, Kyung-Mi;Heo, Moon-Soo
    • Journal of Life Science
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    • v.28 no.8
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    • pp.955-961
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    • 2018
  • Although much research has focused on various bioactive substances in starfish, research on microorganisms isolated from starfish is lacking as compared with other natural products. In this study, we investigated bacterial communities in the gut of red starfish (Certonardoa semiregularis) in Jeju Island. In total, 103 bacterial strains were isolated using marine agar and R2A medium. The isolated strains were analyzed and identified using the 16S rRNA gene sequence. Based on an analysis of this gene sequence, the 103 isolated bacteria were classified into four major groups: Proteobacteria (93%: Alpha-proteobacteria, 24.8%; Beta-proteobacteria, 4%; Gammaproteobacteria, 65%) Bacteroidetes (4%), Actinobacteria (2%), and Firmicutes (1%). In addition, the isolates were divided into seven classes (Actinobacteria, Flavobacteria, Bacilli, Sphingobacteria, Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria), 15 orders, 19 families, and 24 genera. A phylogenetic analysis revealed two strains, Lysobacter sp. and Pedobacter sp., with similarity of 97.55% and 97.58%, respectively. As the similarity in the 16S rRNA gene sequence was 98% or less compared to previously identified bacteria, the two strains may possibly be classified as a new genus or species. We suggest that additional studies, including biochemical and morphological tests, should be performed to identify the new candidate strains.

Bacterial Diversity in the Mud Flat of Sunchon Bay, Chunnam Provice, by 16S rRNA Gene Analysis (16S rRNA 유전자 분석에 의한 전남 순천만 갯벌의 세균 다양성)

  • 이명숙;홍순규;이동훈;배경숙
    • Korean Journal of Microbiology
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    • v.37 no.2
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    • pp.137-144
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    • 2001
  • In order to investigate the diversity of bacterial community in the mud flat of Sunchon Bay, Chunnam province, diversity of amplified 16S rDNA was examined. Total DNA was extracted from sediment soils and 16S rDNAs were amplified using PCR primers based on the universally conserved sequences in bacteria. Clonal libraries were constructed and 111 clones were examined by amplified rDNA restriction analysis (ARDRA) using HaeIII. Clones were clustered based on restriction patterns using computer program, GelCompar II. One hundred different RFLP types were detected from 111 clones. The 20 clones were selected and sequenced according to dendrograms derived from ARDRA, to cover most of the bacterial diversity in the clone libraries. None of the clones were identical to any representatives in the Ribosomal Database Project small subunit RNA databases and GenBank. All sequences showed between 77 and 96.8% similarity to the known 16s rRNA sequence from cultured organisms. The 20 clones sequenced fell into seven major lineages of the domain Bacteria: alpha-, delta-, gamma-Proteobacteria, low G+C Gram positive bacteria, high G+C Gram positive bacteria, Sphingobacteria (Cytophaga) and Cyanobacteria (chloroplast). Among the clones, the Proteobacteria were dominant.

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Using Reverse Dot Hybridization Method and 16S rRNA Gene (16S rDNA) for Identifying the Food Poisoning Microorganism in Foods (Reverse dot hybridization 방법과 16S rRNA gene(16S rDNA)을 이용한 식품에서 식중독균의 탐색)

  • Kim, Min-Seong;Shin, Kyu-Chul;Lee, Hyung-Gu;Han, Myung-Soo;Min, Byung-Re;Choi, Yong-Keel
    • Korean Journal of Food Science and Technology
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    • v.35 no.3
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    • pp.470-474
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    • 2003
  • DNA sequence information on small-subunit rRNA gene (16S rDNA) obtained from food-poisoning bacterial culture was used to investigate the presence of bacterial pathogens in food. By reverse dot blot detection method, presence of food-poisoning bacteria could be confirmed on hybridization of digoxigenin-labeled 16S rDNA Polymerase Chain Reaction (PCR) primer product and biotin-labeled specific oligonucleotide probe. Escherichia coli, Bacillus cereus. and Salmonella sp. were used as the representative food-poisoning bacterial microorganisms. An oligonucleotide probe, based on the variable region of 16S rRNA gene, was used as the specific probe. These tools may be more useful than classic biochemical method for rapid identification of contaminated food.

Phylogenetic Relationships of the Aphyllophorales Inferred from Sequence analysis of Nuclear Small Subunit Ribosomal DNA

  • Kim, Seon-Young;Jung, Hack-Sung
    • Journal of Microbiology
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    • v.38 no.3
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    • pp.122-131
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    • 2000
  • Phylogenetic classification of the Aphyllophorales was conducted based on the analysis of nuclear small subunit ribosomal RNA (nuc SSU rDNA) sequence. Based on phylogenetic groupings and taxonomic characters, 16 families were recognized and discussed. Although many of the characters had more or less homoplasies, miroscopic characters such ad the mitic system and clamp, spore amyloidity and rot type appeared to be important in the classification of the Aphyllophorales. Phylogenetically significant families were newly defined to improve the classification of the order Aphyllophorales.

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Sequence analysis of LSU rDNA of Alexandrium tamarense/catenella complex from Korean coastal waters

  • Kim, Keunyong;Kim, Chang-Hoon
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2001.05a
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    • pp.252-254
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    • 2001
  • A great deal of effort has been put into the identification of Alexandrium tamarense/fundyense/catenella complex by understanding correlation between morphological and subcellular characteristics. To date, the most promising tool for the study of these species is sequence analyses of rRNA genes that have been useful for various organisms' taxonomy and phylogeny, and its application such as in situ hybridization. (omitted)

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Cloning and Characterization of Genes Controlling Flower Color in Pharbitis nil Using AFLP (Amplified Fragment Length Polymorphism) and DDRT (Differential Display Reverse Transcription)

  • Kim, Eun-Mi;Jueson Maeng;Lim, Yong-Pyo;Yoonkang Hur
    • Journal of Photoscience
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    • v.7 no.2
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    • pp.73-78
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    • 2000
  • To analyze molecular traits determining pigmentation between Pharbitis nill violet and white, Amplified Fragment Length Polymorphism(AFLP) and Differential Display Reverse Transcription(DDRT) experiments were carried out with either genomic DNAs or total RNAs isolated from both plants. Results of AFLP experiment in combination of 8 EcoRⅠ primers with 6 MseⅠ primers showed 41 violet-and 60 white-specific DNA bands. In the subsequent experiment, 22 violet-and 22 white-specific DNA fragments were amplified by PCR with DNAs eluted. The sizes of the fragments range from 200 to 600bp. DDRT using total RNA produced 19 violet-and 17 white-specific cDNA fragments, ranging from 200 to 600bp. The fragments obtained by both AFLP and DDRT had been cloned into pGEM T-easy vector, amplified and subjected to the nucleotide sequence analyses. As a result of Blast sequence analysis, most of them sequenced up to date showed no similarity to any Known gene, while few has similarity to known animal or plant genes. An AFLP clone V6, for example, has a strong sequence similarity to the human transcription factor LZIP-alpha mRNA and a DDRT clone W19 to Solanum tuberosum 3-hydroxy-3-methylglutaryl coenzyme A reductase mRNA.

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Genetic Variation of Rice Populations Estimated Using nrDNA ITS Region Sequence

  • Wang, Dong;Hong, Soon-Kwan
    • Korean Journal of Plant Resources
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    • v.27 no.3
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    • pp.249-255
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    • 2014
  • The rice belonging to Oryza sativa is not only has significant economic importance, for it is the major source of nutrition for about 3 billion all around the world. But also plays a vital role as a model organism, because it has a number of advantages to be a model plant, such as efficient transformation system and small genome size. Many methods and techniques have been conducted to attempt to distinguish different Oryza sativa species, such as amplified fragment length polymorphism (AFLP), random amplified polymorphic DNA (RAPD), simple sequence repeat (SSR) and so on. However, studies using sequence analysis of internal transcribed spacer (ITS), a region of ribosomal RNA has not been reported until now. This study was undertaken with an aim to understand the phylogenetic relationships among sixteen isolates of Oryza sativa collected from abroad and fifteen isolates collected from Korea, using ribosomal RNA (rRNA) internal transcribed spacer (ITS) sequences to compare the phylogeny relationships among different Oryza sativa species. The size variation obtained among sequenced nuclear ribosomal DNA (nrDNA) ITS region ranged from 515bp to 1000bp. The highest interspecific genetic distance (GD) was found between Sfejare 45 (FR12) and Anapuruna (FR15). Taebong isolate showed the least dissimilarity of the ITS region sequence with other thirty isolates. This consequence will help us further understanding molecular diversification in intra-species population and their phylogenetic analysis.

Two Groups of Phytoplasma from Chrysanthemum (Dendranthema grandiflorum) Distinguished by Symptoms and 16S rRNA Gene Sequence in Korea

  • Chung, Bong-Nam;Kim, Byung-Dong
    • The Plant Pathology Journal
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    • v.21 no.2
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    • pp.132-136
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    • 2005
  • Two groups of phytoplasma were identified in chrysanthemum(Dendranthema grandiflorum) cv. Chunkwang showing distinct symptoms. Isolate Ph-ch1 showed symptoms of dwarf, witches'-broom, rosette and root death. The other isolate, Ph-ch2, revealed symptoms of dwarf, yellowing, leaf cupping, vein clearing and root death. The presence of phytoplasma structures in chrysanthemum leaf tissue was confirmed by transmission electron microscopy. The 16S rRNA gene was amplified from isolates Ph-ch1 and Ph-ch2 by PCR and cloned, and the nucleotide sequences were determined. In RFLP analysis, isolate Ph-ch2 showed profiles identical to Ph-ch1, except with restriction enzymes HhaI and MseI. The sequence data showed that isolate Ph-ch1 was most closely related to the aster yellows (AY) phytoplasma, and isolate Ph-ch2 was more closely related to stolbur phytoplasma than to AY phytoplasma. This is the first reported observation of stolbur phytoplasma in chrysanthemum species.