• 제목/요약/키워드: 28S rDNA sequence

검색결과 98건 처리시간 0.023초

Rapid Detection of Enterobacter sakazakii Using TaqMan Real-Time PCR Assay

  • Kang, Eun-Sil;Nam, Yong-Suk;Hong, Kwang-Won
    • Journal of Microbiology and Biotechnology
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    • 제17권3호
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    • pp.516-519
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    • 2007
  • Enterobacter sakazakii is an emerging food pathogen, which induces severe meningitis and sepsis in neonates and infants, with a high fatality rate. The disease is generally associated with the ingestion of contaminated infant formula. In this study, we describe the development of a real-time PCR protocol to identify E. sakazakii using a TaqMan probe, predicated on the nucleotide sequence data of the 168 rRNA gene obtained from a variety of pathogens. To detect E. sakazakii, four primer sets and one probe were designed. Five strains of E. sakazakii and 28 non-E. sakazakii bacterial strains were used in order to ensure the accuracy of detection. The PCR protocol successfully identified all of the E. sakazakii strains, whereas the 28 non-E. sakazakii strains were not detected by this method. The detection limits of this method for E. sakazakii cells and purified genomic DNA were 2.3 CFU/ assay and 100 fg/assay, respectively. These findings suggest that our newly developed TaqMan real-time PCR method should prove to be a rapid, sensitive, and quantitative method for the detection of E. sakazakii.

Rapid Origin Determination of the Northern Mauxia Shrimp (Acetes chinensis) Based on Allele Specific Polymerase Chain Reaction of Partial Mitochondrial 16S rRNA Gene

  • Kang, Jung-Ha;Noh, Eun-Soo;Park, Jung-Youn;An, Chel-Min;Choi, Jung-Hwa;Kim, Jin-Koo
    • Asian-Australasian Journal of Animal Sciences
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    • 제28권4호
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    • pp.568-572
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    • 2015
  • Acetes chinensis is an economically important shrimp that belongs to the Sergestidae family; following fermentation, A. chinensis' economic value, however, is low in China, and much of the catch in China is exported to Korea at a low price, thus leading to potential false labeling. For this reason, we developed a simple method to identify A. chinensis' origin using allele-specific polymerase chain reaction (PCR). Ten single nucleotide polymorphisms (SNPs) were identified from partial (i.e., 570 bp) DNA sequence analysis of the mitochondrial 16s rRNA gene in 96 Korean and 96 Chinese individual shrimp. Among 10 SNP sites, four sites were observed in populations from both countries, and two sites located in the middle with SNP sites at their 3'-ends were used to design allele-specific primers. Among the eight internal primers, the C220F primer specific to the Chinese A. chinensis population amplified a DNA fragment of 364 bp only from that population. We were able to identify the A. chinensis population origin with 100% accuracy using multiplex PCR performed with two external primers and C220F primers. These results show that the 16S rRNA gene that is generally used for the identification of species can be used for the identification of the origin within species of A. chinensis, which is an important finding for the fair trade of the species between Korea and China.

Copper Tolerance of Novel Rhodotorula sp. Yeast Isolated from Gold Mining Ore in Gia Lai, Vietnam

  • Kim Cuc Thi Nguyen;Phuc Hung Truong;Cuong Tu Ho;Cong Tuan Le;Khoa Dang Tran;Tien Long Nguyen;Manh Tuan Nguyen;Phu Van Nguyen
    • Mycobiology
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    • 제51권6호
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    • pp.379-387
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    • 2023
  • In this study, twenty-five yeast strains were isolated from soil samples collected in the gold mining ore in Gia Lai, Vietnam. Among them, one isolate named GL1T could highly tolerate Cu2+ up to 10 mM, and the isolates could also grow in a wide range of pH (3-7), and temperature (10-40 ℃). Dried biomass of GL1 was able to remove Cu2+ effectively up to 90.49% with a maximal biosorption capacity of 18.1 mg/g at pH 6, temperature 30 ℃, and incubation time 60 min. Sequence analysis of rDNA indicated this strain was closely related to Rhodotorula mucilaginosa but with 1.53 and 3.46% nucleotide differences in the D1/D2 domain of the 28S rRNA gene and the ITS1-5.8S rRNA gene-ITS2 region sequence, respectively. Based on phylogenetic tree analysis and the biochemical characteristics, the strain appears to be a novel Rhodotorula species, and the name Rhodotorula aurum sp. nov. is proposed. This study provides us with more information about heavy metal-tolerant yeasts and it may produce a new tool for environmental control and metal recovery operations.

Effect of Culture Conditions on Cathepsin B Inhibitor Production by a Marine Bacterium, Pseudomonas sp. Strain PB01

  • Hoang, Le Thu Van;Kim, Moon-Moo;Kim, Se-Kwon
    • Journal of Microbiology and Biotechnology
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    • 제18권6호
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    • pp.1115-1120
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    • 2008
  • A novel cathepsin B inhibitor-producing bacterium was isolated from marine sediments and identified based on its 16S rDNA sequence as Pseudomonas sp. strain PB01 (Accession No. EU126129). The growth and enzyme inhibitor production were investigated under various culture conditions. A mixture of organic nitrogen source was required for the optimal production, whereas both glucose and maltose proved to be the effective carbon sources for cathepsin B inhibitor production. Other optimal culture conditions included temperature range between 25 and $28^{\circ}C$, initial medium pH of 6.6, and shaking speed of 200 rpm. Under these optimal conditions, the maximum inhibitory activity from culture broth was approximately 50% after 30 h of cultivation. Additionally, kinetic study revealed that inhibitor production paralleled with cell growth, which suggested that the inhibitor may be a primary metabolite of that bacterium.

Isolation and Characterization of Three Zygomycetous Fungi in Korea: Backusella circina, Circinella muscae, and Mucor ramosissimus

  • Nguyen, Thuong T.T.;Lee, Hyang Burm
    • Mycobiology
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    • 제46권4호
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    • pp.317-327
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    • 2018
  • While surveying undiscovered fungal taxa in Korea, three rare zygomycetous fungal strains, CNUFC-PTF2-1, CNUFC-TF3-1, and CNUFC-ESAF3-1, were isolated from soil, leaf, and freshwater samples, respectively. The strains were analyzed morphologically as well as phylogenetically based on the internal transcribed spacer region and 28S rDNA sequences. Sequence analysis of the two loci revealed that the isolates, CNUFC-PTF2-1, CNUFC-TF3-1, and CNUFC-ESAF3-1, were identified as Backusella circina, Circinella muscae, and Mucor ramosissimus, respectively. These species have not yet been previously described in Korea.

청자갈치(Bothrocara hollandi)의 근육에 기생하는 점액포자충Myxobolus aeglefini (Myxozoa: Myxobolidae) (Myxobolus aeglefini (Myxozoa: Myxobolidae) infection in muscles of porous-head eelpout (Bothrocara hollandi))

  • 전찬혁;김정호
    • 한국어병학회지
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    • 제28권2호
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    • pp.79-85
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    • 2015
  • 동해안에서 어획된 청자갈치 (Bothrocara hollandi)의 체측 근육에 유백색의 불투명한 시스트를 형성하고 있는 점액포자충이 발견되었다. 시스트를 마쇄하여 광학현미경으로 관찰해본 결과, 원형에 가까운 점액포자충의 성숙 포자가 관찰되었다. 성숙 포자의 평균 길이는 $11.9(11.0{\sim}13.5){\mu}m$, 평균 폭은 $11.6(10.7{\sim}13.6){\mu}m$, 평균 두께는 $7.8(6.9{\sim}8.8){\mu}m$이었다. 극낭의 평균 길이는 $4.4(3.2{\sim}5.3){\mu}m$이었으며, 평균 폭은 평균 $3.3(2.4{\sim}4.2){\mu}m$이었다. 감염숙주와 성숙포자의 형태학적 특징, 각 부위의 측정값으로부터 본 연구에서 발견된 점액포자충은 Myxobolus aeglefini Auerbach 1906으로 동정하였다. 또한 18S rDNA sequences를 이용한 계통분석 결과 M. albi와 M. groenlandicus와 같은 분지에 속하는 것이 확인되었으며 각각 97.7%와 96.9%의 상동성을 나타내었다.

Kiwi-persimmon wine produced using wild Saccharomyces cerevisiae strains with sugar, acid, and alcohol tolerance

  • Hee Yul Lee;Kye Man Cho;Ok Soo Joo
    • 한국식품저장유통학회지
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    • 제30권1호
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    • pp.52-64
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    • 2023
  • 100 different yeast colonies were isolated from spontaneously fermented kiwis, persimmons, apples, pears, watermelons, grapes, grape fruits, peachs, and plums, and selected yeast strains were used to produce kiwi-persimmon mixed wine (KPMW). Among the isolates, five representative strains exhibited tolerance to sucrose, alcohol, pH, and potassium metabisulfite when compared with the control yeast strain (Saccharomyces cerevisiae KCCM 12615). All five yeast strains (Y4, Y10, Y28, Y78, and Y81) exhibited 99% 26S rDNA sequence similarity to S. cerevisiae. The pH, acidity, Brix, reducing sugar, alcohol, and organic acid contents were consistent in KPMW prepared from the S. cerevisiae KCCM 12615 and Y28 strains. KPMW made from the Y4, Y10, and Y28 strains exhibited lower quantities of free sugars than those of the KPMW made from the other yeast strains. The level of ethyl esters in KPMW prepared from the Y28 was higher than that in the other KPMWs. All strains, except for Y28, produced lower concentrations of sulfur and ketone compounds. Furthermore, the KPMW produced by the Y28 strains had total phenolic contents with 1.1 g/L, with DPPH and ABTS radical scavenging activities of 57.06% and 55.62%, respectively, and a FRAP assay value of 0.72. Our results suggest that Y28 is a promising yeast strain for producing high-quality wines.

부영양 호수에서 분리한 Acidovorax temperans AK-05의 Anabaena cylindrica 분해 특성 (Characterization of a Novel Alga-Lytic Bacterium, Acidovorax temperans AK-05, Isolated from an Eutrophic Lake for Degradation of Anabaena cylindrica)

  • 김정동;한명수
    • 생태와환경
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    • 제37권2호통권107호
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    • pp.241-247
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    • 2004
  • 부영양 호수로부터 살조 세균을 분리하고 동정한 결과 Anabaena cylindrica NIES-19를 유릴 탄소원으로 이용하는 double layer방법으로 15종의 살조세균을 분리하였으며 높은 살조 활성을 나타내는 4종의 살조세균 AK-05, AK-07, AK-13 그리고 AK-28을 선별하여 살조 능력을 비교하였다. 이들 중에 AK-05가 가장 높은 살조 활성을 나타내었으며 이를 16S rDNA염기서열을 분석한결과 Acidovorax temperans와 99.5%의 유사성을 나타내어 Acidovorax temperans AK-05로 명명하였다. A. temperans AK-05의 배양 여액을 A. cylindrica NIES-19에 뚜렷한 살조 활성을 나타내었으며, 이것의 살조 활성 능력을 분석한 결과 살조 활성에 관여하는 주요 물질은 non-protein이며 열에 안정적이었다. 이러한 살조 활성 능력은 알칼리 조건과 25${\sim}$$30^{\circ}C$에서 가장 높게 나타냈다. 따라서 이와 같은 특성은 일반적으로 알칼리 조건을 야기하는 Cyanobacteria에 의한 water blooms이 발생하는 호수에 적용하는데 매우 유리할 것으로 여겨진다.

Complete mitochondrial genome of Rotunda rotundapex Miyata & Kishida 1990 (Lepidoptera: Bombycidae), which was named as Bombyx shini Park & Sohn 2002

  • Park, Jeong Sun;Kim, Min Jee;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • 제44권2호
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    • pp.55-64
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    • 2022
  • Bombyx shini Park & Sohn, 2002 (Lepidoptera: Bombycidae), which was listed as an endemic species in South Korea has recently been renamed as the East Asian silk moth Rotunda rotundapex Miyata & Kishida, 1990 (Lepidoptera: Bombycidae). In this study, we sequenced the complete mitochondrial genome (mitogenome) of the R. rotundapex to announce genomic characteristics and to clarify its validity with a new name. The 15,294-bp long complete mitogenome comprises a typical set of genes [13 protein-coding genes (PCGs), 2 rRNA genes, and 22 tRNA genes] and one major noncoding, A + T-rich region, with an arrangement identical to that observed in most lepidopteran mitogenomes. The A/T content of the whole mitogenome was 79.22%; however, it varied among the regions/genes as follows: A + T-rich region, 91.62%; srRNA, 84.67%; lrRNA, 83.01%; tRNAs, 81.43%; and PCGs, 77.46%. Phylogenetic analyses of 35 species in the Bombycoidea superfamily showed the sister relationship between the families Sphingidae and Bombycidae s. str., with the higher nodal support [bootstrap support (BS) = 78%]. The Saturniidae was placed as the sister to the two families, but the nodal support for this relationship was low (BS = 53%). Current R. rotundapex was placed together with previously reported con-species with the highest nodal support, forming a separate clade from Bombyx, validating that B. shini can have a new genus name, Rotunda. However, the Korean R. rotundapex showed a substantial sequence divergence at 5.28% to that originated from an individual of type locality Taiwan in 1,459-bp of COI sequences. Considering such a high sequence divergence an additional study, which includes morphological and DNA barcoding data from further extensive distributional range maybe is needed for further robust taxonomic conclusion.

한국 자생 난초 뿌리에서 분리한 미기록 내생균 5종 (Five Species of Endophytic Fungi Isolated from Roots of Native Orchid Plants from Korea)

  • 이봉형;엄안흠
    • 한국균학회지
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    • 제45권4호
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    • pp.355-361
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    • 2017
  • 본 연구는 전라북도 진도에서 자란(Bletilla striata), 강원도 정선군 함백산에서 감자난초(Oreorchis patens)와 은대난초(Cephalanthera longibracteata)의 뿌리에서 내생균을 분리하고 이들의 형태 및 분자생물학적 방법을 통해 동정하였다. 각 균주의 형태학적 특징을 분석하기 위해 포자의 모양과 특징을 관찰하였고, 분자생물학적 동정을 위해 ITS1F와 ITS4 primer를 이용하여 internal transcribed spacer (ITS) 및large subunit (LSU) rDNA영역을 분석하였다. 그 결과 Phialocephala bamuru, Coniochaeta mutabilis, Phialophora foetens, Calonectria Canadensis, Neonectria ramulariae 5종의 국내 미기록 내생균이 동정되어 이를 보고하고자 한다.