• Title/Summary/Keyword: PCR 동정

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Some Properties of an Isolate of Peanut stunt virus Isolated from White Clover (Trifolium repens L.) (토끼풀에서 분리한 Peanut stunt virus의 성질)

  • Jung, Goo-Ho;Jeon, Yong-Woon;Choi, Jang-Kyung;Hong, Jin-Sung;Ryu, Ki-Hyun;Lee, Sang-Yong
    • Research in Plant Disease
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    • v.14 no.1
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    • pp.71-75
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    • 2008
  • An isolate of Peanut stunt virus (PSV), named as Tr-PSV, was isolated from white clover (Trifolium repens L) showing mosaic symptom. Tr-PSV systemically infected all plants tested in the Nicotiana spp. and induced local lesions on inoculated leaves of Chenopodium amaranticolor. However, Tr-PSV induced typical mosaic symptoms as ER-PSV on Vigna unguiculata 5 to 6 days after inoculation, while Fny-CMV used as a control virus of Cucumovirus produced local lesions on inoculated leaves. In dsRNA analysis, Tr-PSV consisted of four dsRNAs, but satellite RNA was not detected. The cDNA of coat protein gene of Tr-PSV was amplified by RT-PCR using a Cucumovirus-specific single pair primers that designed to amplify a DNA fragment of approximately 950 bp. By restriction mapping analysis using RFLP of the RT-PCR products and by serological properties of gel diffusion test, Tr-PSV belongs to a typical member of PSV subgroup I. This is the first report on the occurrence of PSV in white clover in Korea.

RAPD-SCAR Markers Linked to Medium-Leaf Zoysiagrass Ecotypes (한국잔디 중지 변이개체와 연관된 RAPD-SCAR 마커)

  • Chung, Sung Jin;Park, Su Jeong;Kim, Hun Joong;Yang, Geun-Mo;Choi, Joon-Soo;Oh, Chan-Jin;Jang, Deok-Hwan;Song, In-Ja;Lee, Geung-Joo
    • Weed & Turfgrass Science
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    • v.2 no.2
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    • pp.191-197
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    • 2013
  • Two medium-leaf ecotypes (CY6069, CY6097) belonging to one species (Zoysia japonica) of Korean lawngrasses were selected in sod production fields in Jang Seong, Korea. They were reported to have distinct morphological and growth rate characteristics different from the preferred medium-leaf type zoysiagrass in Korea. This study was conducted to define further the genotypic difference at the molecular level and to develop DNA marker based on the specific DNA fragment. Polymorphic DNA fragments were first explored by using randomly amplified polymorphic DNA (RAPD) primers, which were then converted into PCR-based sequence characterized amplified region (SCAR) markers. The CY6069-specific primer set amplified about 550 bp successfully, while the CY6097 marker produced the expected 690 bp band, by which those markers were nominated by CY6069_550 and CY6069_690 SCARs, respectively. Together with the reported morphological and other phenotypic features, the SCAR markers confirmed in this study will be useful to identify those medium-leaf zoysiagrass genotypes when they are cultivated with other vegetatively propagated warm-season turfgrasses in sod farms.

Density and Distribution of the Mosquito Population Inhabiting Jeju Region, 2018 (제주지역에 서식하는 모기 개체군 밀도와 분포, 2018)

  • Seo, Min Young;Chung, Kyoung A
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.336-343
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    • 2019
  • In order to investigate the density of seasonal incidence of mosquitoes, a vector of infectious diseases in Jeju region, this study collected mosquitoes using a Black light trap (BL) and Biogents' Sentinel 2 Mosquito Trap (BG), dividing the region into cattle sheds, habitats for migratory birds, and the downtown area, twice a month for 9 months from March through November 2018. Then, this study conducted identification and classification and checked for the presence of Flavivirus using reverse transcription polymerase chain reaction (RT-PCR). As for the mosquito population, 1,847 mosquitoes (six genera, 12 species) were collected. The places where most mosquitoes were collected were copses near craft workshops in habitats for migratory birds and Jungang-dong in the Seogwipo downtown area. For the population, Culex pipiens pallens was the dominant species (76.9%), followed by Aedes albopictus (8.9%). Most of the population was collected in June, followed by August and October. This study conducted a RT-PCR test with 1,847 collected mosquitoes, which were divided into 50 pools if they had Flavivirus. All turned out to be negative. However, the results of the investigation show the presence of Culex tritaineniorhychus, Aedes albopictus, and Anopheles sinensis and can be used as a basis for the comprehensive prevention management of mosquitoes.

Pests occurring on Cymbidium (심비디움에 발생하는 해충의 종류)

  • Cho, Myoung Rae;Jeon, Sung-Wook;Kang, Taek Joon;Kim, Hyung Hwan;Ahn, Seung-Joon;Yang, Chang Yeol
    • Korean journal of applied entomology
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    • v.52 no.4
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    • pp.403-408
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    • 2013
  • A survey of pest occurrence and status of farmer's pest management was conducted at 45 cymbidium farms in 10 major cultivation areas in Korea. The pest species collected from the cymbidium farms were identified as follows: Tetranychus urticae Koch, Frankliniella intonsa Trybom, Pinnaspis aspidistrae Signoret, Incilaria confusa Cockarel, Halyomorpha brevis Walker, Myzus persicae S$\ddot{u}$lzer, and Aphis gossypii Glover, Coccus hesperidum Linnaeus, Thrips flavus Schrank, and Thrips tabaci Lindeman. The two-spotted spider mite, T. urticae, was the key pest in cymbidium production, occurring on 45 farms, followed by scales (20 farms), slugs (6), thrips (8), aphids (5), and stinkbug (1). PCR-RFLP of the rDNA ITS2 region revealed that two thrips species, Thrips flavus Schrank and Thrips tabaci Lindeman, occur on cymbidium farms. Therefore, it is necessary for the cymbidium farmers to establish an integrated pest management system to meet quarantine standards.

Identification and characteristics of DDX3 gene in the earthworm, Perionyx excavatus (팔딱이 지렁이(Perionyx excavatus) DDX3 유전자의 동정 및 특성)

  • Park, Sang Gil;Bae, Yoon-Hwan;Park, Soon Cheol
    • Journal of the Korea Organic Resources Recycling Association
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    • v.23 no.1
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    • pp.70-81
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    • 2015
  • Helicases are known to be a proteins that use the chemical energy of NTP binding and hydrolyze to separate the complementary strands of double-stranded nucleic acids to single-stranded nucleic acids. They participate in various cellular metabolism in many organisms. DEAD-box proteins are ATP-dependent RNA helicase that participate in all biochemical steps involving RNA. DEAD-box3 (DDX3) gene is belonging to the DEAD-box family and plays an important role in germ cell development in many organisms including not only vertebrate, but also invertebrate during asexual and sexual reproduction and participates in stem cell differentiation during regeneration. In this study, in order to identify and characterize DDX3 gene in the earthworm, Perionyx excavatus having a powerful regeneration capacity, total RNA was isolated from adult head containing clitellum. Full length of DDX3 gene from P. excavatus, Pe-DDX3, was identified by RT-PCR using the total RNA from head as a template. Pe-DDX3 encoded a putative protein of 607 amino acids and it also has the nine conserved motifs of DEAD-box family, which is characteristic of DEAD-box protein family. It was confirmed that Pe-DDX3 has the nine conserved motifs by the comparison of entire amino acids sequence of Pe-DDX3 with other species of different taxa. Phylogenetic analysis revealed that Pe-DDX3 belongs to a DDX3 (PL10) subgroup of DEAD-box protein family. And it displayed a high homology with PL10a, b from P. dumerilii.

Altering of Collagens in Early Pregnant Mouse Uterus (착상전 생쥐 자궁에서 콜라겐의 변화)

  • Cheon, Yong-Pil
    • Development and Reproduction
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    • v.11 no.1
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    • pp.1-11
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    • 2007
  • Specific endometrial preparation should occur during periimplantation period. That is a progress of serial differentiation and is absolute in implantation of embryo and successful pregnancy. Remodeling of tissues shown during embryogenesis is regulated by various factors including extracellular matrix (ECM). Marked changes during pregnancy are including embryo migration, decidual response, and differentiation of placenta in placental animals including human. These changes to successful implantation in embryo and uterus have to prepare the competence for attachment of embryo and uterus, and invasion defense of uterus. During these changes, ECM dramatically changes for maintaining the uterine and embryonic functions. The major component of most connective tissue is collagens. It is very complex and hard to explore the mechanisms for ECM modulation. Recently using high throughput methodology, PCR-select cDNA subtraction method, microarray, many candidate genes have been identified. Steroid hormones have fundamental role in implantation and maintenance of pregnancy. Dermatopontin, a regulator of collagen accumulation, is regulated spatio-temporally in the uterus by primarily progesterone through progesterone receptors at the time of implantation. Modulation of extracellular matrix is critically regulated by cascade of gene net-works which are regulated by cascade of sex steroid hormones. Pathological regulation of uterine extracellular matrix reported in diabetic patients. To know the extracellular modulation is essential to understanding implantation, feto-placental development and overcome the paths involved in female reproduction. Though ECM composed with very various components and it is complex, the present review focused on the fate of collagens during periimplantation period.

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Identification of Daphne Mottle Virus Isolated from Daphne odora, a New Member of the Genus Potyvirus (서향에서 분리한 신종 포티바이러스(Daphne Mottle Virus)의 동정)

  • Park, Chung Youl;Park, Jungan;Lee, Boo-Ja;Bak, Sangmin;Lee, Hong-Kyu;Kim, Jeong-Sun;Yoon, Youngnam;Suh, Sang Jae;Lee, Su-Heon
    • Research in Plant Disease
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    • v.22 no.1
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    • pp.59-63
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    • 2016
  • A new poty-like virus was isolated from plants of winter daphne (Daphne odora) that showed virus-like symptoms on leaves, from four regions of Korea during 2014. Filamentous-shaped particles were observed by transmission electron microscopy of preparations extracted from symptomatic leaves and examined by the direct negative stain method. RT-PCR assay showed that three samples were positive for both Cucumber mosaic virus and potyvirus, and only one sample was positive for potyvirus only. A BLAST comparison to partial sequences from helper-component proteinase, cylindrical inclusion and coat protein genes detected the highest nucleotide identity of 76%, 72%, and 72% with Daphne mosaic virus, respectively, levels below the potyvirus species discrimination threshold. The new potyvirus was isolated using indicator plants (Chenopodium amaranticolor), in which local lesions were produced. In this study, we identified a novel potyvirus from winter daphne, which we have named Daphne mottle virus (DapMoV).

Expression of Sex-Related Genes in the Fetus of Mouse: 2-Bromopropane and Sex Differentiation (생쥐 태자의 성 관련 유전자 발현: 2-Bromopropane과 성 분화)

  • Choi, Donchan;Lim, Sinae;Kim, Pan Gyi;Kim, Dae-Yong;Lee, Young-Soon
    • Development and Reproduction
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    • v.5 no.2
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    • pp.107-114
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    • 2001
  • The recent reports that endocrine disruptors(EDs) bring about abnormalities in reproductive organs and functions of invertebrates suggest that mammals be affected by the EDs. The present study examined the influence of 2-bromopropane(2-BP) by looking at the sexes of litters in mouse. The expression of sex-related genes during sex differentiation was also investigated in the fetus of mouse. The male and female mice were infused with 2-BP for 3 weeks before mating. The litters were sexed at the weaning time from the 4 different groups. The sex-related genes were identified by RT-PCR from the fetuses at gestation 10 days. The sequences of the genes were analysed by comparing to those of other animals. The mean numbers of litters survived by the weaning time were slightly reduced in the only group of both female and male mice treated with 2-BP. The female litters were greater than male litters in the only group of female treated with 2-BP. The other groups showed male litters greater than female litters. The sex-related genes, SRY, DAX1, SF1 , and AMH genes were identified and sequenced, showing 416, 466, 326, 389 base pairs, respectively. All of the genes had the homology of 89~90% with rat and 81~92% with human within the range of bases identified. They were expressed at the time of sex determination. Therefore, it appears that 2-BP somewhat affects the reproductive activity of adult mouse. Influence of 2-BP on the reproductive function is expected to be studied through the expression of the sex-related genes.

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Early Identification of Citrus Zygotic Seedlings Using Pollen-specific Molecular Markers (화분 특이적 마커를 이용한 감귤 교잡종 실생묘의 조기 동정)

  • Jin, Seong Beom;Yun, Su Hyun;Park, Jae Ho;Park, Suk Man;Koh, Sang Wook;Lee, Dong Hoon
    • Horticultural Science & Technology
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    • v.33 no.4
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    • pp.598-604
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    • 2015
  • This study was carried out to develop molecular techniques to allow the selection of zygotic seedlings in the early stage of the plant development. We identified 37 pollen-specific molecular markers from RAPD analysis and successfully used them for identification of the zygotic seedlings from various hybrid crosses. Three Satsuma mandarin cultivars ('Morita unshiu', 'Nangan 20' and 'Miyagawawase') were used as mother parents and seven cultivars ('Ponkan', 'Lee', 'Kinokuni', 'Shiranuhi', 'Tamnaneunbong', 'Shinyegam', and 'Sunburst' mandarins) served as pollen parents. PCR analysis showed that 2 primers could identify zygotic hybrid seedlings. Among them, an UBC-27 primer was used to identify the zygotic seedlings from hybrid crosses of "'Nangan 20' ${\times}$ 'Kinokuni'" mandarin, "'Nangan 20 ${\times}$ Ponkan'" mandarin and "'Miyagawawase ${\times}$ Sunburst'" tangerine. In total 29 out of 40 seedlings (73%), 9 out of 47 seedlings (19%), and 13 out of 45 (29%) were identified as zygotic seedlings, respectively. These results can show that the pollen-specific markers selected in this study can be used effectively for early identification of zygotic seedlings from Citrus hybrid crosses.

A Pathotype of Pepper mild mottle virus Causing Necrotic Spot Symptoms in Paprika Fruit (파프리카 과실에 괴사반점을 일으키는 Pepper mild mottle virus의 병원형)

  • Choi, Gug-Seoun;Choi, Sung-Kook;Cho, Jeom-Deog;Cho, In-Sook
    • Research in Plant Disease
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    • v.19 no.2
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    • pp.124-127
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    • 2013
  • Black necrotic spots were observed from the fruits of paprika that were cultivating in a vinylhouse. The casual agents of the symptom were identified as several isolates of Pepper mild mottle virus (PMMoV) by responses of indicator plants, electron microscopy, and RT-PCR analysis. Symptoms of the viral disease were mild mottle in the young leaves, necrotic spots on the fruits and the fruit apex of paprika, but the symptoms were not shown on the mature leaves. All of the PMMoV isolates were determined as $P_{1.2.3}$ pathotypes from the biological responses on the chilli pepper lines used for discrimination of tobamovirus pathotypes. Pathogenicity of the PMMoV isolates was also confirmed using mechanical inoculation method to paprika seedlings. The coat protein (CP) genes of the PMMoV isolates were compared at the nucleotide and amino acid levels with the previously published PMMoV isolate. The isolates share 96 to 99% CP nucleotide identity among the isolates. The CP of $P_{1.2}$-pathotype PMMoV-P2 presented Met at position 139, But the CPs of $P_{1.2.3}$-pathotype PMMoVs from paprika showed Met to Asn substitution at the same position. This is the first report of identification of $P_{1.2.3}$-pathotype PMMoV isolates from paprika in Korea.