• 제목/요약/키워드: ITS1-5.8S-ITS2

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체외배양 기술로 생산된 초기배에 의한 한우 송아지 생사기술 개발. II. 체외성숙, 수정된 소 초기배의 체외발생에 있어서 ITS와 EGF의 효과

  • 서경덕;김호중;김갑수;김광식
    • 한국가축번식학회지
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    • 제22권1호
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    • pp.51-59
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    • 1998
  • This study was undertaken to investigate the effects of ITS and EGF on embryonic development of in vitro matured and fertilized bovine oocytes in culture medium su, pp.ementing with or without calf serum. When fertilized oocytes were cultured in TCM-199 containing 0, 0.1, 0.5 and 1.0\ulcorner/ml ITS with 5% calf serum, the rates of development to blastocyst stage and the cell number of blastocysts were not significantly different among all treatments. And also monolayer of cumulus/granulosa cells prepared in containing calf serum and ITS were no beneficial effects of embryonic development. On the other hand, when EGF was su, pp.imented to TCM-199 containing calf serum or calf serum free, embryonic development rates(24.0 2.8% to 29.2 1.7% or 8% to 9%) and cell number of blastocysts(p<0.05) were significantly increased compared with EGF-free(22.1 2.1 or 1.0%, p<0.05). But when fertilized oocytes were cultured with cumulus/granurosa cells in TCM-199 containing EGF and calf serum, the rate of embryos development to the blastocyst stage and cell number of blastocysts were not significantly different compared with EGF-free and any concentrations. These results showed that ITS and EGF was not improved the development of bovine embryo in vitro matured and in vitro fertilized with calf serum and/or monolayer of cumulus/granulosa cells.

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DNA 바코딩과 고해상 융해곡선분석에 기반한 인삼속 식물의 종 판별 (Internal Transcribed Spacer Barcoding DNA Region Coupled with High Resolution Melting Analysis for Authentication of Panax Species)

  • 방경환;김영창;임지영;김장욱;이정우;김동휘;김기홍;조익현
    • 한국약용작물학회지
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    • 제23권6호
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    • pp.439-445
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    • 2015
  • Background : Correct identification of Panax species is important to ensure food quality, safety, authenticity and health for consumers. This paper describes a high resolution melting (HRM) analysis based method using internal transcribed spacer (ITS) and 5.8S ribosomal DNA barcoding regions as target (Bar-HRM) to obtain barcoding information for the major Panax species and to identify the origin of ginseng plant. Methods and Results : A PCR-based approach, Bar-HRM was developed to discriminate among Panax species. In this study, the ITS1, ITS2, and 5.8S rDNA genes were targeted for testing, since these have been identified as suitable genes for use in the identification of Panax species. The HRM analysis generated cluster patterns that were specific and sensitive enough to detect small sequence differences among the tested Panax species. Conclusion : The results of this study show that the HRM curve analysis of the ITS regions and 5.8S rDNA sequences is a simple, quick, and reproducible method. It can simultaneously identify three Panax species and screen for variants. Thus, ITS1HRM and 5.8SHRM primer sets can be used to distinguish among Panax species.

Morphological and molecular characterization of the genus Coolia (Dinophyceae) from Bahía de La Paz, southwest Gulf of California

  • Morquecho, Lourdes;Garate-Lizarraga, Ismael;Gu, Haifeng
    • ALGAE
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    • 제37권3호
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    • pp.185-204
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    • 2022
  • The genus Coolia A. Meunier 1919 has a global distribution and is a common member of epiphytic dinoflagellate assemblages in neritic ecosystems. Coolia monotis is the type species of the genus and was the only known species for 76 years. Over the past few decades, molecular characterization has unveiled two species complexes that group morphologically very similar species, so their limits are often unclear. To provide new knowledge on the biogeography and species composition of the genus Coolia, 16 strains were isolated from Bahía de La Paz, Gulf of California. The species were identified by applying morphological and molecular approaches. The morphometric characteristics of all isolated Coolia species were consistent with the original taxa descriptions. Phylogenetic analyses (large subunit [LSU] rDNA D1 / D2 and internal transcribed spacer [ITS] 1 / 5.8S / ITS2) revealed a species assemblage comprising Coolia malayensis, C. palmyrensis, C. tropicalis, and the C. cf. canariensis lineage. This is the first report of Coolia palmyrensis and C. cf. canariensis in Mexico and C. tropicalis in the Gulf of California. Our results strengthen the biogeographical understanding of these potentially harmful epiphytic dinoflagellate species.

Degradation of Malic Acid by Issatchenkia orientalis KMBL 5774, an Acidophilic Yeast Strain Isolated from Korean Grape Wine Pomace

  • Seo, Sung-Hee;Rhee, Chang-Ho;Park, Heui-Dong
    • Journal of Microbiology
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    • 제45권6호
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    • pp.521-527
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    • 2007
  • Several yeast strains degrading malic acid as a sole carbon and energy source were isolated from Korean wine pomace after enrichment culture in the presence of malic acid. Among them, the strain designated as KMBL 5774 showed the highest malic acid degrading ability. It was identified as Issatchenkia orientalis based on its morphological and physiological characteristics as well as the nucleotide sequences of the internal transcribed spacer (ITS) 1-5.8S rDNA-ITS II region. Phylogenetic analysis of the ITS I-5.8S rDNA-ITS II sequences showed that the KMBL 5774 is the closest to I. orientalis zhuan 192. Identity of the sequences of the KMBL 5774 was 99.5% with those of I. orientalis zhuan 192. The optimal pH of the media for the growth and malic acid degradation by the yeast was between 2.0 and 3.0, suggesting that the strain is an acidophile. Under the optimized conditions, the yeast could degrade 95.5% of the malic acid after 24 h of incubation at $30^{\circ}C$ in YNB media containing 2% malic acid as a sole carbon and energy source.

Study of antimicrobial activity and the mode of action of Anal P5 peptide

  • Park, Yoonkyung;Hahm, Kyung-Soo
    • 통합자연과학논문집
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    • 제1권1호
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    • pp.47-53
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    • 2008
  • In a previous study, we showed that Cecropin A (1-8)-Magainin 2 (1-12) hybrid peptide (CA-MA)'s analogue, Anal P5, exhibit broad-spectrum antimicrobial activity. Anal P5, designed by flexible region (positions 9, 10)-substitution, Lys- (positions 4, 8, 14, 15) and Leu- (positions 5, 6, 12, 13, 16, 17, 20) substitutions, showed an enhanced antimicrobial and antitumor activity without hemolysis. The primary objective of the present study was to gain insight into the relevant mechanisms of antimicrobial activities of Anal P5 by using flow cytometric analysis. Anal P5 exhibits strong antifungal activity in a salt concentration independent manner. In addition, Anal P5 causes significant morphological alterations of the bacterial surfaces as shown by scanning electron microscopy, supporting its antibacterial activity. Its potent antibiotic activity suggests that Anal P5 is an excellent candidate as a lead compound for the development of novel antibiotic agents.

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밀양근교에서 채집한 야생 동충하초 계통의 PCR 산물에 근거한 계통 유전학적 연구 (Phylogenetic Analysis on Wild Cordyceps Collected from Miryang Region of South Korea)

  • 박현철;이상몽;박남숙
    • 한국자원식물학회지
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    • 제34권1호
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    • pp.1-16
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    • 2021
  • 남부 지방의 밀양 근교에 자생하고 있는 야생 동충하초 균주(Cordyceps sp., Paecilomyces sp., Beauveria sp., Aranthomyces sp., Isaria sp., Himenostilbe sp.)를 채집 분리하여 rDNA 및 internal transcribed spacer (ITS) 부위의 염기서열을 비교하였다. ITS 영역에 특정적인 프라이머인 ITS1과 ITS4를 이용하여 PCR을 수행하여 증폭하였다. 다양한 균주에서 같은 크기의 PCR 생산물을 얻을 수 있었고, 이들의 서열분석을 위하여 pGEM-T easy 벡터에 클로닝하였으며, ITS1, 5.8S, ITS2 영역 부위의 염기서열들을 BLAST 수행하여 유연관계를 분석하였다. 밀양 근교에서 분리한 32개의 균주 중에 Cordyceps militaris는 서열이 등록된 유전정보 AY49191, EU825999, AY491992와 100% 일치하였으며, 몇 개의 종은 보고된 서열과 모두 일치하지는 않았다. 예를 들어 strain P17은 울주군 가지산에서 분리한 P. tenuipes로서 밀양시 조천읍 가지산에서 분리한 P. tenuipes와는 서열상에서 다른 부분들이 있었다. 결론적으로 밀양 근교의 균주들을 분리하는데 ITS영역분석이 분류와 검정에 효율적이고, 본 연구를 통하여 동충하초의 생태학적인 유전자원들을 확보할 수 있었다.

Characterization of Sclerotinia sclerotiorum Isolated from Paprika

  • Jeon, Young-Jae;Kwon, Hyuk-Woo;Nam, Ji-Sun;Kim, Seong-Hwan
    • Mycobiology
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    • 제34권3호
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    • pp.154-157
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    • 2006
  • A fungal isolate collected from infected paprika (Capsicum annuum var. grossum) was characterized as Sclerotinia sclerotiorum based on its ability of sclerotium formation, physiological and molecular properties. When the isolate was grown on potato dextrose agar, oatmeal agar, and malt extract agar, it grew most well on PDA. Optimal temperature and pH for its growth were $25^{\circ}C$ and pH 7, respectively. The fungal isolate produced sclerotia on PDA within 10 days, and the color and shape of the sclerotia were similar to those of S. sclerotiorum. The ITS rDNA regions including ITS1 and ITS2 and 5.8S sequences were amplified using ITS1F and ITS4 primers from the genomic DNAs of the paprika isolate and other known pathogenic S. sclerotiorum isolated from different crops in Korea, and their nucleotide sequences were determined. Sequence comparison analysis showed the ITS rDNA of the paprika isolate shares 100% sequence identity with those of S. sclerotiorum isolated from red pepper, lettuce and a S. sclerotiorum isolate registered in GenBank DNA database. Neighbor joining analysis based on the ITS rDNA sequence revealed the paprika isolate has very close phylogenetic relationships with known Sclerotinia sclerotiorum isolates. This is the first report that S. sclerotiorum has been found associated with paprika rot in paprika growing countries.

신두리 해안사구에 자생하는 사구식물 내생진균의 다양성 분석 (Endophytic Fungal Diversity Associated with the Roots of Coastal Sand-dune Plants in the Sindu-ri Coastal Sand Dune, Korea)

  • 유영현;서영교;윤혁준;김현;김예은;이리나 할무라토바;임순옥;김창무;김종국
    • 한국미생물·생명공학회지
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    • 제41권3호
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    • pp.300-310
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    • 2013
  • 98주의 내생진균은 신두리 해안사구에 자생하고 있는 해안사구식물의 뿌리로부터 분리하였다. 8종의 해안사구식물 샘플은 모래지치(Argusia sibirica), 갯메꽃(Calystegia soldanella), 갯그령(Elymus mollis), 반디지치(Lithospermum zollingeri), 갯무(Raphanus sativus), 솔장다리(Salsola collina), 왕잔디(Zoysia macrostachya) 및 갯잔디(Zoysia sinica)이며, 신두리 해안사구로부터 채집되었다. 그리고 분리된 내생진균들은 ITS1, 5.8S와 ITS2를 포함하는 ITS-rDNA 영역에 의해 분석되었다. 해안사구식물로부터 분리된 내생진균에 대하여 다양한 지수를 적용하여 분석하였다. 해안사구식물로부터 분리된 모든 내생진균은 Capnodiales (3.09%), Eurotiales (70.10%), Glomerellales (1.03%), Helotiales (3.09%), Hypocreales (9.28%), Mortierellales (2.06%), Onygenales (1.03%), Ophiostomatales (1.03%), Pleosporales (1.03%), Polyporales (1.03%), Russulales (1.03%), Saccharomycetales (2.06%), Xylariales (1.03%)로 13개 목과 분류체계가 명확하지 않은 Incertae sedis (3.09%)으로 확인되었다. 본 연구에서는 8종의 식물로부터 내생진균을 분석한 결과 Eurotiales 목과 Hypocreales 목의 Penicillium 속(59.18%)과 Fusarium속 (5.10%)이 가장 많이 분포하는 것으로 나타났다. 갯메꽃으로부터 분리된 내생진균이 다른 해안사구식물들로부터 분리된 내생진균의 다양성 보다 높은 것으로 확인되었다.

Ribosomal DNA ITS 유전자를 이용한 왕지렁이(빈모강: 지렁이과) 그룹의 계통분류 (Molecular Phylogeny of the Amynthas-complex (Oligochaeta: Megascolecidae) Inferred from ITS Nucleotide Sequences)

  • 홍용;;황의욱;이보은;박순철;김태흥
    • 환경생물
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    • 제25권4호
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    • pp.349-355
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    • 2007
  • 지렁이과 (family Megascolecidae) 왕지렁이속 (genus Amynthas) 그룹을 rDNA ITS 유전자를 이용하여 계통 유연관계를 알아보고자 하였다. 2과 10속에서 26종의 DNA 염기서열을 이용하여 종간 계통적 유연관계를 MP (Maximum Parsimony), NJ (Neighbor Joining), QP (Quartet Puzzling)로 계통도를 작성하였다. 염기서열이 확보된 Megascolecidae (지렁이과)의 국내외 26종에 대한 계통 분석은 위 3개의 알고리즘 모두 동일한 5개의 그룹으로 나뉘는 것을 확인하였다. 또한 Amynthas 속은 크게 한국산 그룹과 필리핀산 그룹으로 구분되었다. 첫 번째 그룹, Amynthas jirensis, A. agrestis, A. gucheonensis, A. sopaikensis, A. bubonis, A. multimaculatus, A. koreanus, A. dageletensis, A. heteropodus, A. odaesanensis, Pontoscolex sp., Pheretima sp. 1와 다른 그룹, Dendropheretima banahawensis. Amynthas halconensis, A. isarogensis, A. mindrooensis, Pithemera sp. 2, Pithmera sp. 1, and Pleionogaster sp.이다. 이는 ITS 유전자가 속간의 차이를 보여주는 것보다 동일한 속이라 하더라도 지리적 차이에 따른 환경의 영향을 받았음을 알 수 있다. 즉 형태적 분화가 이루어졌다고 하더라도 유전적으로는 동질성을 나타내고 있음을 알 수 있다. 형태적 특징에 의한 명백한 차이는 구분되지 않았다.