• Title/Summary/Keyword: RFLP.

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Molecular Identification of Meloidogyne spp. in Soils from Fruit and Vegetable Greenhouses in Korea (분자기법을 이용한 과채류 시설재배지 토양 내 분포하는 뿌리혹선충의 종 동정)

  • Kim, Se-Jong;Yu, Yong-Man;Whang, Kyung-Sook
    • Korean journal of applied entomology
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    • v.53 no.1
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    • pp.85-91
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    • 2014
  • In this study, we analyzed the phylogenetic characterization of root-knot nematodes (Meloidogyne spp.) in soils from fruits and vegetables greenhouses in Korea. Soil samples were collected from 12 greenhouse fields in which tomato, cucumber, watermelon, and Oriental melon were being cultivated. Meloidogyne spp. were detected in all the soil samples at an average number of $72{\pm}6$ nematodes/300 g of soil to $2,898{\pm}468$ nematodes/300 g of soil. Phylogenetic analysis using polymerase chain reaction-restriction fragment length polymorphism was attempted for the second-stage juveniles (J2) of Meloidogyne spp. collected from the greenhouse soils. Twelve Meloidogyne spp. from the greenhouse soils were classified into two groups by using HinfI digestion of mitochondrial DNA, resulting in 900, 410, 290, and 170 bp fragments (group A) and 900, 700, and 170 bp fragments (group B). Phylogenetic analysis based on mitochondrial DNA sequences (1,483-1,521 bp) showed that nine group A isolates were identified as Meloidogyne incognita (99.73-99.93%) and three group B isolates showed 99.54-99.73% similarity to Meloidogyne arenaria.

DNA Markers Applicable for Identification of Two Internal Apple Feeders, Grapholita molesta and Carposina sasakii (두 종의 사과 심식나방류 [복숭아순나방 (Grapholita molesta), 복숭아심식나방 (Carposina sasakii)] 동정용 DNA 분자지표)

  • Song, Seung-Baeck;Choi, Kyeung-Hee;Lee, Soon-Won;Kim, Yong-Gyun
    • Korean journal of applied entomology
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    • v.46 no.2
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    • pp.175-182
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    • 2007
  • Two fruit moths of the oriental fruit moth, Grapholita molesta (Busck), and the peach fruit moth, Carposina sasakii (Matsumura), infest apples in Korea by internally feeding behavior. C. sasakii is a quarantine insect pest from some other countries importing Korean apples. G. molesta is not a quarantine insect pest, but can be incorrectly identified as C. sasakii especially when it is found inside apple fruits at its larval stages because it is not easy to identify the two species by morphological characters alone. This incomplete identification results in massive economical loss by fruits needlessly destroyed or turned away at border inspection stations of the importing nations. This difficulty can be overcome by molecular DNA markers. Several polymorphic regions of mitochondrial DNA of both species were sequenced and used for developing specific striction sites and polymerase chain reaction (PCR) primers. Based on these sequences, three diagnostic PCR-restriction fragment length polymorphism (RFLP) sites were detected and validated for their practical uses. Also, species-specific PCR primers were devised to develop diagnostic PCR method for identifying the internal feeders.

ACTIVATION OF H-RAS ONCOGENE IN RAT SALIVARY GLAND TUMORS INDUCED BY DMBA AND IRRADIATION (DMBA 매식과 방사선 조사로 유도된 백서 타액선 종양에서 H-ras 암유전자의 활성화)

  • Hu Key-Soon;Choi Jong-Whan;Choi Soon-Chul;Park Tae-Won;You Dong-Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.28 no.1
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    • pp.245-259
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    • 1998
  • Cellular transforming genes have been identified in a number of different tumor cell lines and tumor types. A significant number of these oncogenes belong to the ras gene family. The ras gene family consists of three closely related genes:H-ras, K-ras and N-ras which code for a related 21 kDa protein. Mutations in codon 12, 13 and 61 of one of the three ras genes convert these genes into acute oncogenes. The presence of H-ras gene mutations has important prognostic implications in various tumors. Each genomic DNA was isolated from tumors induced by implantation with DMBA, or by treatment with DMBA -implantation/irradiation. When genome DNA was transfected into NIH 3T3 cells and investigated by two-step PCR-RFLP, the fOllowing results were concluded: 1. Transformation foci developed in two groups when the genome DNA of two experimental groups were transfected into NIH 3T3 cells. 2. Transformation efficiency was 0.01-0.02 foci/㎍DNA in the experimental group with the DMBA-implantation, 0.01-0.03 foci/㎍lgDNA in the experimental group with the DMBA-implantation/irradiation according to results of transfection assay. 3. When the point mutation of H-ras gene was investigated by a two-step PCR-RFLP, there was 13.9% (5/36) in the experimental group with the DMBA implantation, 15.4 % (6/39) in the experimental group with the DMBA -implantation/irradiation. 4. The point mutation in codon 12 and 61 of H-ras was 5.6%(2/36) and 8.3%(3/36) in the experimental group with the DMBA implantation. 5. The point mutation in codon 12 and 61 of H-ras gene was 7.7%(3/39) in the experimental group with the DMBA -implantation/irradiation.

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Biogeochemical Reactions in Hyporheic Zone as an Ecological Hotspot in Natural Streams (자연 하천의 생태학적 중요 지점으로서 지표수-지하수 혼합대의 생지화학적 기작)

  • Kim, Young-Joo;Kang, Ho-Jeong
    • Journal of Wetlands Research
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    • v.11 no.1
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    • pp.123-130
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    • 2009
  • Hyporheic zone is an area where hydraulic exchanges occur between surface water and ground water. Such transient area is anticipated to facilitate diverse biogeochemical reactions by providing habitats for various microorganism. However, only a few data are available about microbial properties in hyporheic zone, which would be important in better understanding of biogeochemical reactions in whole streams. The study site is Naesung stream, located in the north Kyoung-Sang Province, of which sediment is sandy with little anthropogenic impacts. Soil samples were collected from a transect placed perpendicular to stream flow. The transect includes upland fringe area dominated by Phragmites japonica, bare soil, and soil adjacent to water. In addition, soil samples were also collected from downwelling and upwelling areas in hyporheic zone within the main channel. Soils were collected from 3 depth in each area, and water content, pH, and DOC were measured. Various microbial properties including extracellular enzyme activities ($\beta$-glucosidase, N-acetylglucosaminidase, phosphatase and arylsulfatase), and microbial community structure using T-RFLP were also determined. The results exhibited a positive correlation between water content and DOC, and between extracellular enzyme activities and DOC. Distinctive patterns were observed in soils adjacent to water and hyporheic zone compared with other soils. Overall results of study provided basic information about microbial properties of hyporheic zone, which appeared to be discernable from other locations in the stream corridor.

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Molecular Ecological Characterization of Wastewater Bacterial Communities in Response to Algal Growth (조류성장에 따른 하수 박테리아 군집 변화에 관한 분자생태학적 연구)

  • Lee, Ju-Youn;Lee, Jang-Ho;Park, Joon-Hong
    • Journal of Korean Society of Environmental Engineers
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    • v.33 no.11
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    • pp.847-854
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    • 2011
  • To deal with issues from global climate changes, renewable bioenergy has become important. Algae have been regarded as a good resource for biorefinery and bioenergy, and also have potential capability to remove nutrient and non-decompositional pollutants for wastewater advanced treatment. Although algal-bacterial ecological interaction would be a crucially important factor in using algae for wastewater advanced treatment and resource recovery from wastewater, very little is known about ecological interaction between algae and bacteria in a real wastewater environment. In this study, under a real municipal wastewater condition, we characterized wastewater pollutant treatability and bacterial communities in response to growth of Ankistrodesmus gracilis SAG278-2, which can grow in wastewater and has a high lipid contents. The growth of algal population using the wastewater was inhibited by increase in wastewater bacteria while bacterial survival and cellular decay rate were not influenced by the algal growth. Removals of recalcitrant organic matters and total nitrogen were improved in the presence of algal growth. According to T-RFLP and statistical analysis, algal growth affected time-course changes in bacterial community structures. The following 16S rRNA gene amplicon, cloning results showed that the algal growth changes in bacterial community structure, and that bacterial populations belonging to Sediminibacterium, Sphingobacterium, Mucilaginibacter genera were identified as cooperative with the algal growth in the wastewater.

Characteristics and Improving Breed of Economic Traits of Korea Native Chicken (한국 재래 닭 품종 특성 및 초기성장 개량을 위한 분자표지 개발)

  • Oh J. D.;Park M. H.;Kong H. S.;Lee H. K.;Jeon G. J.;Yeon S. H.;Sang B. D.;Choi C. H.;Cho B. W.
    • Korean Journal of Poultry Science
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    • v.32 no.1
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    • pp.29-34
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    • 2005
  • This study was conducted to estimate the effects of genotype for chicken major histocompatibility complex (MHC) B-LB genes on economic traits. To detect polymorphism, 400 bp fragments of MHC B-LB genes were obtained and sequenced. After digestions using restriction enzyme Hea III, two restriction enzyme sites were observed. There were two mutations at position 427 and 651 those were decided as Type I and Type II, respectively. Using RFLP analyses, type I were genotyped to TT, TC and CC, and type II to MM, Mm and mm. The relatively higher TC genotype frequencies (0.8) of Type I and Mm genotype frequencies (0.88) of Type II were observed in Korean native chickens. The effects of the genotype on 150 days body weight trait were investigated by the associations of CC and Mm genotypes (P<0.05) in Korean native chickens. This result suggests that a significant association exists between the SNP and 150 days body weight.

Identification of Quantitative Trait Loci for Resistance to Soybean Cyst Nematode Race 14 (콩시스트 선충 race14에 대한 저항성 유전자좌 구명)

  • Choi, In-Soo;Kim, Yong-Chul
    • Journal of Life Science
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    • v.13 no.4
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    • pp.375-382
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    • 2003
  • A direct and precise explanation of soybean resistance to soybean cyst nematode will be possible only when the individual gene(s) involved in the resistance are tagged. This study was conducted, (1) to identify and localize quantitative trait loci for resistance to soybean cyst nematode race 14 on RAPD map, (2) to identify the magnitude and mode of inheritance for each quantitative trait loci, and (3) to identify the best combinations of quantitative trait loci for resistance to soybean cyst nematode race 14. Thirty markers (29 RAPD and 1 RFLP) showed significant association with resistance to soybean cyst nematode race 14. From MAPMAKER/QTL analysis, we identified two regions (linkage group C-7 and linkage group C-9) for resistance to soybean cyst nematode .ace 14. The first quantitative trait loci that was localized at 6.0 cM from $H06^1$ on linkage group C-7 showed a dominant inheritance mode. However, we can not exclude the possibility of additive inheritance mode. The second quantitative trait loci that was localized between $B15^2$ and $E01^1$ on linkage group C-9 also showed a dominant mode of inheritance. One pair of flanking markers ($H06^1$ and $H06^2$) and B15$^2$ were used for multiple regression analysis. Marker combination that included 2 markers, $B15^2$ and $H06^1$, explained the highest total variance (22.9%) for resistance to soybean cyst nematode race 14. Further localization of genes for resistance to soybean cyst nematode race 14 and examination of interaction between quantitative trait loci will accelerate the exploitation of resistance to soybean cyst nematode.

Current status of anisakid nematode larvae infection in marine fishes caught from the coastal area of Korea between 2010 and 2012 (2010~2012년 연안에서 서식하는 해산어에서 아니사키스 유충의 감염현황)

  • Kim, Wi-Sik;Jeon, Chan-Hyeok;Kim, Jeong-Ho;Kim, Do-Hyung;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.25 no.3
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    • pp.189-197
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    • 2012
  • A survey was conducted to investigate infection of anisakid nematode larvae in 243 wild marine fishes caught from the southern coastal area of Korea between 2010 and 2012. The samples comprised fishes from 9 orders, 30 families and 50 species. Total infection rate of anisakid nematode larvae was 10.7% (26/243 fish), which comprised from Yeosu, 7.4% (7/95) in 2010 and 22.7% (5/22) in 2011; from Jeju, 8.2% (5/61) in 2011; from Wando, 40.9% (9/22) in 2012. Anisakid nematode larvae were not detected in Tongyoung and Wando samples in 2011. Molecular identification of the 89 worms from 26 fish was conducted by PCR-RFLP and/or sequence analysis of internal transcribed spacer (ITS) region of ribosomal DNA. From the results, 6 kinds of anisakis species were identified: Anisakis pegreffii (infection rate: 53.9%, 48/89 worms), Hysterothylacium aduncum (38.2%, 34/89), H. fabri (3.4%, 3/89), hybird (A. simplex X A. pegreffii) (2.4%, 2/89), A. simplex (1.1%, 1/89) and Raphidascaris lophii (1.1%, 1/89). The rate of single infection was 80.8% (21/26 infected fish), while 19.2% (5/26) showed mixed infection with 2 to 3 different anisakis species.

Molecular Monitoring of Plankton Diversity in the Seonakdong River and Along the Coast of Namhae (분자 모니터링을 이용한 서낙동강과 남해 연안 플랑크톤 군집 분석)

  • Kim, Bo-Kyung;Lee, Sang-Rae;Lee, Jin-Ae;Chung, Ik-Kyo
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.15 no.1
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    • pp.25-35
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    • 2010
  • The biodiversity of eukaryotic plankton has commonly been used to evaluate the status of aquatic ecosystems. Therefore, an accurate and rapid method for species identification is needed to reveal the biodiversity of environmental water samples. To date, molecular methods have provided a great deal of information that has enabled identification of the hidden biodiversity in environmental samples. In this study, we utilized environmental polymerase chain reaction (PCR) and constructed the 18S nuclear ribosomal RNA clone library from environmental water samples in order to develop more efficient methods for species identification. For the molecular analysis, water samples were collected from the Seonakdong River (Gimhae Bridge) and the coast of Namhae,(Namhaedo). Colony PCR and restriction fragment length polymorphism of PCR (PCR-RFLP) were then adopted to isolate unique clones from the 18S rDNA clone library. Restriction fragment length polymorphism pattern analysis of the Gimhae Bridge sample revealed 44 unique clones from a total of 60 randomly selected clones, while analysis of the Namhae sample revealed 27 unique clones from 150 clones selected at random. A BLAST search and subsequent phylogenetic analysis conducted using the sequences of these clones revealed hidden biodiversity containing a wide range of taxonomic groups (Heterokontophyta (7), Ciliophora (23), Dinophyta (1), Chytridiomycota (1), Rotifera (1) and Arthropoda (11) in the Gimhae Bridge samples Ciliophora (4), Dinophyta (3), Cryptophyta (1), Arthropoda (19) in the Namhae samples). Therefore, the molecular monitoring method developed here can provide additional information regarding the biodiversity and community structure of eukaryotic plankton in environmental samples and helps construct a useful database of biodiversity for aquatic ecosystems.

Hypotensive Effect of Scrophularia buergeriana and Gene Analysis of Scrophularia Species (현삼(玄參)의 유전자 분석 및 자발성 고혈압흰쥐의 혈압에 미치는 영향)

  • Park, Kyu-Ha;Ham, In-Hye;Choi, Ho-Young
    • The Korea Journal of Herbology
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    • v.23 no.2
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    • pp.213-223
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    • 2008
  • Objectives : This study was performed to prove the hypotensive effect of 70% MeOH extracts from Scrophularia buergeriana (SB) on rat's femoral artery. Methods : We observed hypotensive effect of SB on rat's femoral artery. Scrophularia buergeriana Miquel, S. kakudensis, S. koraiensis and S. ningpoenesis Hemsl. were used by the RFLP method and sequence analysis of ITS regions, in order to compare and discriminate. Results : SB 70% MeOH Ex., Water fraction, 40% MeOH fraction, 100% MeOH fraction didn't show significantly hypotensive effect. 20% fraction of SB 70% MeOH Ex. showed significantly hypotensive effect. The size of ITS region on Scrophularia sp. is 589${\sim}$590bp and amount of G+C is 59.3% to 59.7%, and it showed the similar of ITS parts on Scrophularia sp. Conclusions : We concluded that SB Ex. have hypotensive activity, and it's more efficient when separated. And Scrophularia sp. showed the similar of DNA analysis.

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