• 제목/요약/키워드: rDNA sequencing

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A new species and a new record of Meghimatium Slugs (Pulmonata: Philomycidae) in Korea

  • Park, Gab-Man
    • 환경생물
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    • 제39권3호
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    • pp.399-405
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    • 2021
  • Until now, five species (Meghimatium bilineatum, M. fruhstorferi, Limax flavus, L. marginatus, Deroceras reticulatum) in three genera of families Philomycidae and Limacidae have been reported in Korea. Philomycidae is a family of air-breathing land snails. Meghimatium hongdoensis sp. nov. is described based on its middle size (60-80 mm in body length), its body coloration (dark-red yellow), no dorsal with streaks, its genitalia, and 16rDNA sequence analysis. Specimens of this new species were collected from Hongdo island, Sinan-gun, Jeollanam-do, Korea. This species is only known from the type locality at Hongdo to date. It could be found in high-humidity sites. Meghimatium uniforme (Laidlaw 1937) was the first one reported in Korea. It was also collected from Gageodo island, Sinan-gun, Jeollanam-do, Korea. In this study, morphological characteristics including their radula and genital structures of these two species were described. Preliminary results of mitochondrial 16S rDNA sequencing and phylogenetic analyses indicated that these species belonged to the Korean clade.

Diversity Analysis of Diazotrophic Bacteria Associated with the Roots of Tea (Camellia sinensis (L.) O. Kuntze)

  • Arvind, Gulati;Sood, Swati;Rahi, Praveen;Thakur, Rishu;Chauhan, Sunita;Nee Chadha, Isha Chawla
    • Journal of Microbiology and Biotechnology
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    • 제21권6호
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    • pp.545-555
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    • 2011
  • The diversity elucidation by amplified ribosomal DNA restriction analysis and 16S rDNA sequencing of 96 associative diazotrophs, isolated from the feeder roots of tea on enriched nitrogen-free semisolid media, revealed the predominance of Gram-positive over Gram-negative bacteria within the Kangra valley in Himachal Pradesh, India. The Gram-positive bacteria observed belong to two taxonomic groupings; Firmicutes, including the genera Bacillus and Paenibacillus; and Actinobacteria, represented by the genus Microbacterium. The Gram-negative bacteria included ${\alpha}$-Proteobacteria genera Brevundimonas, Rhizobium, and Mesorhizobium; ${\gamma}$-Proteobacteria genera Pseudomonas and Stenotrophomonas; and ${\beta}$-Proteobacteria genera Azospira, Burkholderia, Delftia, Herbaspirillum and Ralstonia. The low level of similarity of two isolates, with the type strains Paenibacillus xinjiangensis and Mesorhizobium albiziae, suggests the possibility of raising species novum. The bacterial strains of different phylogenetic groups exhibited distinct carbon-source utilization patterns and fatty acid methyl ester profiles. The strains differed in their nitrogenase activities with relatively high activity seen in the Gramnegative strains exhibiting the highest similarity to Azospira oryzae, Delftia lacustris and Herbaspirillum huttiense.

Identification of Free-Living Amoebas in Tap Water of Buildings with Storage Tanks in Korea

  • Lee, Da-In;Park, Sung Hee;Baek, Jong Hwan;Yoon, Jee Won;Jin, Soo Im;Han, Kwang Eon;Yu, Hak Sun
    • Parasites, Hosts and Diseases
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    • 제58권2호
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    • pp.191-194
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    • 2020
  • Free-living amoebas (FLAs) can cause severe disease in humans and animals when they become infected. However, there are no accurate survey reports on the prevalence of FLAs in Korea. In this study, we collected 163 tap water samples from buildings, apartments, and restrooms of highway service areas in 7 Korean provinces with high population density. All these buildings and facilities have water storage tanks in common. The survey was separated into categories of buildings, apartments, and highway service areas. Five hundred milliliters of tap water from each building was collected and filtered with 0.2 ㎛ pore filter paper. The filters were incubated in agar plates with heated E. coli at 25℃. After axenization, genomic DNA was collected from each FLA, and species classification was performed using partial 18S-rDNA PCR-sequencing analysis. We found that 12.9% of tap water from buildings with storage tanks in Korea was contaminated with FLAs. The highway service areas had the highest contamination rate at 33.3%. All of the FLAs, except one, were genetically similar to Vermamoeba vermiformis (Hartmannella vermiformis). The remaining FLA (KFA21) was very similar to Acanthamoeba lugdunensis (KA/E26). Although cases of human infection by V. vermiformis are very rare, we must pay attention to the fact that one-third of tap water supplies in highway service areas have been contaminated.

Genetic Variation and Species Identification of Thai Boesenbergia (Zingiberaceae) Analyzed by Chloroplast DNA Polymorphism

  • Techaprasan, Jiranan;Ngamriabsakul, Chatchai;Klinbunga, Sirawut;Chusacultanachai, Sudsanguan;Jenjittikul, Thaya
    • BMB Reports
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    • 제39권4호
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    • pp.361-370
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    • 2006
  • Genetic variation and molecular phylogeny of 22 taxa representing 14 extant species and 3 unidentified taxa of Boesenbergia in Thailand and four outgroup species (Cornukaempferia aurantiflora, Hedychium biflorum, Kaempferia parviflora, and Scaphochlamys rubescens) were examined by sequencing of 3 chloroplast (cp) DNA regions (matK, psbA-trnH and petA-psbJ). Low interspecific genetic divergence (0.25-1.74%) were observed in these investigated taxa. The 50% majority-rule consensus tree constructed from combined chloroplast DNA sequences allocated Boesenbergia in this study into 3 different groups. Using psbA-1F/psbA-3R primers, an insertion of 491 bp was observed in B. petiolata. Restriction analysis of the amplicon (380-410 bp) from the remaining species with Rsa I further differentiated Boesenbergia to 2 groupings; I (B. basispicata, B. longiflora, B. longipes, B. plicata, B. pulcherrima, B. tenuispicata, B. thorelii, B. xiphostachya, Boesenbergia sp.1 and Boesenbergia sp.3; phylogenetic clade A) that possesses a Rsa I restriction site and II (B. curtisii, B. regalis, B. rotunda and Boesenbergia sp.2; phylogenetic clade B and B. siamensis; phylogenetic clade C) that lacks a restriction site of Rsa I. Single nucleotide polymorphism (SNP) and indels found can be unambiguously applied to authenticate specie-origin of all investigated samples and revealed that Boesenbergia sp.1, Boesenbergia sp.2 and B. pulcherrima (Mahidol University, Kanchanaburi), B. cf. pulcherrima1 (Prachuap Khiri Khan) and B. cf. pulcherrima2 (Thong Pha Phum, Kanchanaburi) are B. plicata, B. rotunda and B. pulcherrima, respectively. In addition, molecular data also suggested that Boesenbergia sp.3 should be further differentiated from B. longiflora and regarded as a newly unidentified Boesenbergia species.

Identification of 1,531 cSNPs from Full-length Enriched cDNA Libraries of the Korean Native Pig Using in Silico Analysis

  • Oh, Youn-Shin;Nguyen, Dinh Truong;Park, Kwang-Ha;Dirisala, Vijaya R.;Choi, Ho-Jun;Park, Chan-Kyu
    • Genomics & Informatics
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    • 제7권2호
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    • pp.65-84
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    • 2009
  • Sequences from the clones of full-length enriched cDNA libraries serve as valuable resources for functional genomics related studies, genome annotation and SNP discovery. We analyzed 7,392 high-quality chromatograms (Phred value ${\geq}$30) obtained from sequencing the 5' ends of clones derived from full-length enriched cDNA libraries of Korean native pigs including brainstem, liver, cerebellum, neocortex and spleen libraries. In addition, 50,000 EST sequence trace files obtained from GenBank were combined with our sequences to identify cSNPs in silico. The process generated 11,324 contigs, of which 2,895 contigs contained at least one SNP and among them 610 contigs had a minimum of one sequence from Korean native pigs. Of 610 contigs, we randomly selected 262 contigs and performed in silico analysis for the identification of cSNPs. From the results, we identified 1,531 putative coding single nucleotide polymorphisms (cSNPs) and the SNP detection frequency was one SNP per 465 bp. A large-scale sequencing result of clones from full-length enriched cDNA libraries and identified cSNPs will serve as a useful resource to functional genomics related projects such as a pig HapMap project in the near future.

An assessment of the taxonomic reliability of DNA barcode sequences in publicly available databases

  • Jin, Soyeong;Kim, Kwang Young;Kim, Min-Seok;Park, Chungoo
    • ALGAE
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    • 제35권3호
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    • pp.293-301
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    • 2020
  • The applications of DNA barcoding have a wide range of uses, such as in taxonomic studies to help elucidate cryptic species and phylogenetic relationships and analyzing environmental samples for biodiversity monitoring and conservation assessments of species. After obtaining the DNA barcode sequences, sequence similarity-based homology analysis is commonly used. This means that the obtained barcode sequences are compared to the DNA barcode reference databases. This bioinformatic analysis necessarily implies that the overall quantity and quality of the reference databases must be stringently monitored to not have an adverse impact on the accuracy of species identification. With the development of next-generation sequencing techniques, a noticeably large number of DNA barcode sequences have been produced and are stored in online databases, but their degree of validity, accuracy, and reliability have not been extensively investigated. In this study, we investigated the extent to which the amount and types of erroneous barcode sequences were deposited in publicly accessible databases. Over 4.1 million sequences were investigated in three largescale DNA barcode databases (NCBI GenBank, Barcode of Life Data System [BOLD], and Protist Ribosomal Reference database [PR2]) for four major DNA barcodes (cytochrome c oxidase subunit 1 [COI], internal transcribed spacer [ITS], ribulose bisphosphate carboxylase large chain [rbcL], and 18S ribosomal RNA [18S rRNA]); approximately 2% of erroneous barcode sequences were found and their taxonomic distributions were uneven. Consequently, our present findings provide compelling evidence of data quality problems along with insufficient and unreliable annotation of taxonomic data in DNA barcode databases. Therefore, we suggest that if ambiguous taxa are presented during barcoding analysis, further validation with other DNA barcode loci or morphological characters should be mandated.

rDNA-ITS 및 CAPS 분석에 의한 꽃송이버섯 (Sparassis crispa) 수집균주의 계통분류학적 특성구분 (Phylogenetic relationships of medicinal mushroom Sparassis crispa strains using the rDNA-ITS and CAPS analysis)

  • 정종천;이명철;전창성;이찬중;신평균
    • 한국버섯학회지
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    • 제8권1호
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    • pp.27-32
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    • 2010
  • 본 시험은 국내외에서 수집한 꽃송이버섯균 22균주에 대하여 분자생물학적 유연관계를 분석하고자 하였다. 수집균주의 ribosomal DNA의 ITS 영역에 대한 cleaved amplified polymorphic sequence (CAPS) 분석 결과, KACC50866은 다른 균주들과 20%이하의 유연관계를 나타내었으며 나머지 균주들은 90% 이상의 유연관계를 보이면서 4그룹으로 구분되었다. 따라서 이들의 세분화된 분자생물학적 구분을 위하여 rDNA ITS 영역의 염기서열분석을 하여 구분하여 본 결과 KACC50866 균주는 다른 꽃송이버섯균과 유연관계가 매우 낮은 것으로 나타났다. 그리고 나머지 21개 균주는 같은 그룹으로 구분되어 있어 같은 종으로 생각할 수 있으나, 이들을 좀더 세분하기 위해서는 미토콘드리아의 유전자 서열 분석 등이 병행되어야 할 것으로 판단된다.

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스마트팜 재배 병풀의 triterpenes 정량 및 각질형성세포 활성화 효과 (Quantification of triterpenes in Centella asiatica cultivated in a smart farm, and their effect on keratinocyte activation)

  • 박진홍;조성민;이다희;박영민;장환봉;강태진;이기만
    • 한국식품저장유통학회지
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    • 제30권3호
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    • pp.483-491
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    • 2023
  • 본 연구에서는 제주도에서 자생한 병풀을 수집해 스마트팜과 노지에서 재배하고 이를 이용하여 주요성분 및 각질형성세포 활성화에 미치는 영향을 확인 및 비교하였다. 스마트팜 재배 병풀과 노지 재배 병풀의 유전자 확인을 통한 종 분석을 위해, 핵 속의 ITS DNA와 엽록체의 psbA-H DNA를 증폭하여 염기서열을 분석한 후 NCBI 유전자 은행에서 보고된 식물들의 DNA와 비교하였다. 스마트팜 재배 병풀과 노지 재배 병풀의 ITS DNA 염기서열은 유전자 은행의 MH768338.1번 Centella asiatica와 일치하고 엽록체 psbA-H DNA 또한 유전자 은행의 JQ425422.1번 C. asiatica와 일치하였다. 스마트팜 재배 병풀추출물(SEE)과 노지 재배 병풀추출물(FEE)의 triterpene은 HPLC에 의해 분석되었으며, SEE의 madecassoside, asiaticoside, madecassic acid, asiatic acid 함량은 각각 59.31±0.94 mg/g, 46.38±2.26 mg/g, 6.21±1.47 mg/g, 7.04±1.93 mg/g으로 분석되었다. 반면, FEE는 각각 24.38±1.31 mg/g, 21.28±1.44 mg/g, 3.11±1.05 mg/g, 5.40±1.26 mg/g으로 측정되어 SEE가 FEE보다 더 높은 triterpene을 갖는 것이 확인되었다. 사람 각질형성세포에 대한 SEE와 FEE의 독성은 실험된 농도 내에서 관찰되지 않았으며, 스크래치가 유발된 세포 내 회복은 SEE가 FEE보다 더 높은 회복능을 보였다. 따라서, 본 실험 결과 triterpene 함량이 더 높은 스마트팜 재배 병풀이 건강기능식품 소재로서 더 효과적이라고 판단된다.

Rapid Identification of Lactobacillus plantarium in Kimchi Using Polymerase Chain Reaction

  • Kim, Tae-Woon;Min, Sung-Gi;Choi, Dong-Hun;Jo, Jae-Sun;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • 제10권6호
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    • pp.881-884
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    • 2000
  • A polymerase chain reaction (PCR) was performed to rapidly identify Lactobacillus plantarum from type strains and kimchi samples. The PCR experiments were carried out using specific oligonucleotide primer sets based on the 16S rRNA gene sequences of L. plantarum. The expected DNA amplificate of 419 bp was obtained when either purified DNA or whole cells of L. plantarum strains reacted with LP primers, yet not with any of the other strains. The PCR product was confirmed by DNA sequencing. Accordingly, since the PCR method used is simple, specific, and rapid, it will be useful for monitoring and evaluation L. plantarum in the mixed microbial population found in kimchi.

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어육장의 미생물학적 특성 (Characterization of Microorganisms in Eoyukjang)

  • 오유진;오미화;이종미;조미숙;오상석
    • 한국식품과학회지
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    • 제40권6호
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    • pp.656-660
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    • 2008
  • 예로부터 전통 장류는 먹거리로 많이 이용되어 왔으며, 최근에는 장류의 다양한 영양 성분 및 생리 활성 능력이 밝혀지면서 관심이 증대되고 있다. 본 연구에서는 전통 방식으로 제조된 어육장의 발효에 관여하는 미생물을 숙성 기간별로 분석하고 이를 통해 안전성을 평가하였으며, 이와 함께 국내에서 시판되고 있는 간장의 미생물을 분석하여 어육장의 미생물분석과 비교하였다. 어육장은 2개월 단위로 샘플링하여 분석하였으며, 분리된 미생물은 API kit와 16S rDNA sequencing을 이용하여 동정하였다. 어육장의 발효에 전반적으로 관여하는 미생물은 Bacillus 균류와 Saccharomyces cerevisiae였으며, 숙성 초기에는 Aspergillus flavus가 발효에 관여하는 것으로 확인되었다. 어육장은 최종적으로 섭취 전에 달이는 과정을 거치게 되는데 실험결과 달인 후에는 미생물이 존재하지 않았다. 현재 국내에서 시판되고 간장의 경우 총 균수가 0-42 CFU/mL였고, 동정결과 B. subtilis, B. licheniformis, B. pumilus로 확인되었다. 분리된 Bacillus 균류에 대하여 독소여부를 분석한 결과 어육장 및 시판 장류에서 검출된 Bacillus균류는 본 실험에서 분석된 3가지 설사형 독소와 1종의 구토형 독소를 생성하지 않는 균주들인 것으로 확인되었다.