• Title/Summary/Keyword: ms DNA

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Genetic Analysis of Asian Chum Salmon Populations Based on Microsatellite DNA Variation

  • Yoon, Moon-Geun;Abe, Syuiti;Jin, Deuk-Hee
    • Fisheries and Aquatic Sciences
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    • v.10 no.4
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    • pp.186-190
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    • 2007
  • We examined the genetic variability of Asian chum salmon (Oncorhynchus keta) populations using nuclear microsatellite (ms) DNA analysis with four polymorphic loci (OKM4, OKM5, OKM7, and OKM8) in 397 individuals from nine populations, including one in Korea, seven in Japan, and one in Russia. The msDNA gene diversity was highest in the Japanese populations, suggesting greater genetic variation in the populations in Japan than in populations in Korea and Russia. The pairwise $F_{ST}$ estimates based on our msDNA data showed that the Korean population was genetically different from the Japanese and Russian populations, and there were higher $F_{ST}$ estimates between Hokkaido and Honshu populations than between other population pairs. A neighbor-joining tree showed that the Korean population was distinct from two other clusters, representing the populations in Honshu and the populations in Hokkaido and Russia. These results suggest that the observed population genetic patterns of Asian chum salmon might be influenced by low or restricted gene flow.

Cloning and Characterization of Homeodomain-Zip Gene, Phc5 in Embryogenic Callus derived from Pimpinella brachycarpa Suspension Cultured Cells (참나물 현탁배양세포 유래 배발생캘러스에서 HD-Zip 유전자, Phc5의 클로닝과 특성)

  • 손수인;김준철
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.2
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    • pp.121-126
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    • 1999
  • Calli were induced from the petiole explants of Pimpinella brachycarpa on MS medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L BA after four weeks of culture. Compact clusters of small and dense cells among these calli were selected and suspension-cultured as the source of embryogenic calli. When transferred to MS medium with 0.1 mg/L NAA, the suspension-cultured cells grew to embryogenic callus. Somatic embryos derived from these embryogenic calli developed into plantlets. The cDNA library was constructed in the embryogenic callus and in order to screen the cDNA library, these cDNAs were plated at a density 1.5 $\times$ 10^5 plaques per 15 cm petridish. Among 19 clones showing preferential hybridization with petiole HD-Zip gene, five clones were obtained after second screening. Four clones among them, were highly homologous to P. brachycarpa shoot-tip Phz4 gene, but one clone, Phc5 was about 1.5 kb which has an extra 163 bp to 5' upstream of Phz4. The Phc5 was 1,531 bp containing poly A tails of 18 bases. ATG start codon for Phc5, was located at position 284 with an open reading frame of 906 by which encodes a polypeptide of 302 amino acids. The Phc5 protein revealed that the polypeptides between 135 and 195 contain a homeodomain as the `leucine zipper' motif.

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Genetic Relationships of Sandfish (Arctoscopus japonicas) from Five Different Areas of Korea and Japan Based on Mitochondrial DNA and Microsatellite Analyses (Mitochondrial DNA와 microsatellite marker 분석을 통한 한국과 일본에 서식하는 5 지역의 도루묵(Arctoscopus japonicas)에 대한 유전학적 유연관계 분석)

  • Kim, Eun-Mi;Kang, Hyun-Sook;Kang, Jung-Ha;Kim, Dong-Gyun;An, Cheul Min;Lee, Hae Won;Park, Jung Youn
    • Journal of Life Science
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    • v.25 no.11
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    • pp.1204-1213
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    • 2015
  • A comprehensive analysis of the population structure of the sandfish (Arctoscopus japonicas), the most abundant fishery resource in the East Sea of Korea, has not been carried out, despite its importance in Korea. The present study examined the genetic diversity and differences between five populations (two Japanese and three Korean populations) of A. japonicas captured in the East Sea using both the 401 bp sequence of mitochondrial DNA (mtDNA, cytochrome b) and five microsatellite DNA (msDNA) markers. The results of the analysis using the Cyt b sequence revealed 27 haplotypes. Based on msDNA variations, the estimated expected heterozygosity (HE) in each population ranged from 0.68 (Gampo, Korea) to 0.7765 (Erimo, Japan). Pairwise FST and AMOVA tests using both the Cyt b sequence and msDNA data pointed to significant differences between the Korean and Japanese populations (mtDNA; FST=0.2648, p<0.05, msDNA; FST=0.0814, p<0.05). These results were similar to the results of UPGMA, PCA, and structure analysis. In these analyses, the five populations were assigned to two groups (Korean populations and Japanese populations). These results shed light on the genetic diversity and relationships of A. japonicas and contribute to research on the evaluation, conservation, and utilization of Korean A. japonicas as genetic resources.

Establishment of Genetic Characteristics and Individual Identification System Using Microsatellite loci in Domestic Beef Cattle (초위성체 DNA표지인자를 이용한 국내 육우집단의 품종특성 및 개체식별 체계설정)

  • Kim, Sang-Wook;Jang, Hee-Kyung;Kim, Kwan-Suk;Kim, Jong-Joo;Jeon, Jin-Tae;Yoon, Du-Hak;Kang, Seong-Ho;Jung, Hyo-Il;Cheong, Il-Cheong
    • Journal of Animal Science and Technology
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    • v.51 no.4
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    • pp.273-282
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    • 2009
  • DNA marker information is used to identify or distinguish cattle breeds or individual animal. The purpose of this study was to apply Bovine Genotypes Kit Version 1.1/2.1 to bovine DNA samples (National Institute of Animal Science) taken from Australian / American beef (n=148), Holstein beef (n=170) and Hanwoo cattle (n=177) bred in Jeongeub, Jeonbuk, Korea, so that it could distinguish Hanwoo breed. The Bovine Genotype Kits consist of 16 ISAG MS markers, which were used to build a database of genotypes in each group. Genotyping results were analyzed using MS Tool kit and Phylip program to create phylogenetic tree. The GeneClass 2.0 was used to estimate breed identification. These analyses found that this kit had 100% capacity to distinguish Hanwoo beef, 95.3% capacity to differentiate Australian / American beef and 90% capacity to identify Korean Holstein steer beef. Hence, it is expected that 16 commercial microsatellite markers is useful to categorizegenetic characteristics of Hanwoo breed and also identify Hanwoo individuals and the origin of beef. In particular, it is expected that these markers will be advantageous in discriminating domestic Holstein beef from Australian / Americanbeef.

Non-invasive Biological Monitoring of DNA Adducts Formed at Workers Handling 3,3-Dichlorobenzidine(DCB) by Using GC/MS

  • Lee, Jin-Heon
    • Journal of Environmental Health Sciences
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    • v.29 no.4
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    • pp.21-26
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    • 2003
  • We examine the metabolites(DCB and acetyl DCB) extracted from exfoliated urothelial cells of 33 workers who employed DCB-handling industries. The characteristics of workers submitted urine, whose age, working years and smoking persons were 41.9$\pm$11.1, 8.7$\pm$5.5 and 25(32.0%), respectively. DNA adduct was isolated from the exfoliated urothelial cells by applying $^{32}$ p-postlabeling procedure. Metabolites(DCB and acetyl DCB) were extracted from DNA adducts by hydrolyzing and N-glycosylase. Concentrations of DCB and acetyl DCB were 28.6$\pm$5.25 ng/g DNA, and 17.0$\pm$3.73 ng/g DNA, respectively. The regression between DCB level and exposure years of workers is y = 1.668 + 2.588x(p = 0.005, $r^2$= 0.394). The regression between acetyl DCB level and exposure years of workers is y = 8.071 + 1.325x(p = 0.076, $r^2$= 0.222). Smoking workers are significantly higher than non-smoking workers on DCB and acetyl DCB level(p = 0.065 and 0.021, respectively). DCB level was 33.9$\pm$7.14 ng/g DNA on smokers, and 23.1$\pm$9.97 ng/g DNA on non-smokers. Acetyl DCB was 25.1$\pm$5.27 ng/g DNA on smokers, and 8.92$\pm$7.22 ng/g DNA on non-smokers.

Expression of Canavalia Iineata Leghemoglobin cDNA in Transgenic Nicotiana tabacum (형질전환된 담배에서 해녀콩 Leghemoglobin cDNA의 발현)

  • 이선영
    • Journal of Plant Biology
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    • v.38 no.2
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    • pp.203-209
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    • 1995
  • Tobacco (Nicotiana tahacum L. cv. Wisconsin 38) leaf discs were cocultivated with Agrohacterium carrying a leghemoglobin (Lb) cDNA from Canavalia lineata. Seven plants were regenerated from the transformed leaf discs on MS media supplemented with 0.5 mg/L BAP, 0.1 mg/L ${\alpha}-NAA$, 200 mg/L kanamycin and 500 mg/L carbenicillin. Southern hybridization and PCR of genomic DNA from transgenic plants showed that the Lb cDNA was stably integrated into the genome of the tobacco. Total RNA from the transgenic tobacco showed northern hybridization signal at 1,000 nt and PCR of the first strand cDNA synthesized from the total RNA amplified 0.5 kb Lb cDNA. Furthermore, western hybridization using a polyclonal antibody against soybean Lb showed a 15.8 kD LB-like band on SDS-PAGE of proteins from the transformed tobacco. These results demonstrated that the Lb cDNA of C. lineata was not only incorporated into the genome of tobacco, but also transcribed into mRNA and translated into Lb protein in the transformed tabacco.

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The Introduction of Proteinase Inhibitor II (PI-II) Gene into Flowering Cabbage, Brassica oleracea var. acephala DC. (꽃양배추로의 Proteinase Inhibitor II ( PI-II ) 유전자 도입)

  • 김창길;정재동;안진흥
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.1
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    • pp.45-50
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    • 1998
  • Hypocotyl explants of flowering cabbage were precultured on MS medium without kanamycin and then cocultured with Agrobacterium tumefaciens LBA4404;;pGA875 harboring insect resistantce proteinase inhibitor II(PI-II) gene in MS liquid medium adjusted pH 5.5 for 72hr. These explants were transferred to MS medium containing 20 mg/L kanamycin, 500 mg/L carbenicillin, and 1 mg/L BA. The explants were subsequently subcultured every 2 weeks. After 4 weeks of subculture, kanamycin-resistant shoots were obtained from selection medium. Leaves of putative transformants survived on MS selection medium containing 30 mg/L kanamycin. Incoporation of the PI-II gene into flowering cabbage was confirmed by PCR analysis of genomic DNA. Southern blot analysis showed that ECL-labeled probe for PI-II gene was hybridized to the expected amplified genomic DNA fragment of about 500 by from transgenic flowering cabbage.

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Induction of Somatic Embryos and RAPD Analysis in Regenerated Plantlets of Bupleurum falcatum L. (자호(紫胡)의 체세포배(體細胞胚) 형성(形成)과 재생(再生) 식물체(植物體)의 RAPD 분석(分析))

  • Park, Cheol-Ho;Yu, Chang-Yeon;Seo, Jeong-Sik;Kim, Ki-Sik;Ahn, Sang-Duek;Chang, Byoung-Ho
    • Korean Journal of Medicinal Crop Science
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    • v.3 no.1
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    • pp.50-55
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    • 1995
  • This study was conducted to determine the optimum conditions for induction and different somatic of somatic embryos as well as germination of encapsulated and stored somatic embryos. Somatic embryos was better formed in 1/2X MS medium than full - strength MS medium. 0.1 to 1.0mg/lBA and kinetin promoted shoot differentiation of somatic embryos. Higher concentration tend to inhibit differentiation. IAA affect positively both root and shoot growth. In vitro germination of somatic embryos encapsulated with 2% alginate matrix containing 1/2 MS nutrient medium and $AgNO_3$ 5mg/l was 86%. Storage of somatic embryos was effecive at $5^{\circ}C$ but the germination rate decreased with longer storage period. RAPD analyses with plants regenerated from the somatic embryos showed DNA polymorphism, indicating abolition of primer binding site by point mutation, deletion, or insertion of certain sequences.

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A Study on the Speciadon of a Fresh Water Fish Zacco temminckL VII. Vadation of Mitochondrial DNA between 2 Types of Zacco temmincki (갈겨니(Zacco temminki)의 진화에 관한 연구 VII.갈겨니 2 Type의 Mitochondrial DNA변이)

  • Lee, Hei-Yung;Yang, Suh-Yung;Paik, Sang-Gi;Park, Chang-Shin;Yu, Sung-Lim;Lee, Sung-Keun
    • The Korean Journal of Zoology
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    • v.31 no.3
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    • pp.236-242
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    • 1988
  • Mitochondrial DNAs of two Mdh allelotypes of the dark chub, Z. temmincki inhabiting in Korean fresh water, were analysed. Samples of each type were collected from four populations, and the fragment patterns for mtdNA of each type were explained from 7 of the eleven restriction enzymes with hexanucleotide recognition site. Genome size was approximately 16.7 kilobases. The highly typical mtdNA fragments of each type were discovered in digestion profiles produced by Eco RI and Pst I enzyrnes. The comparisons of restriction fragment patterns and relative digestion maps permitted the estimation of fragment homology (F) and nucleotide sequence divergence(p). Between the two identical types, sequence divergence(p) was 0.128(MS), and 0.045(MM), ; between the two different types, 0.195 (range 0.177-0.226). These result may provide a distinct difference more than the value derived from allozyrne analysis, and a powerful new molecular approach for assessing genetic-evolutionary relationship among fishes.

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A Study on 10 Metabolites Separated from DNA Adduce of Blood Lymphocytes in Rats Exposed Orally with 3,3-dichlorobenzidine(DCB) by GC/MS-SIM

  • Shin, Ueon-Sang;Lee, Jin-Heon
    • Journal of Environmental Health Sciences
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    • v.28 no.4
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    • pp.6-11
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    • 2002
  • 3.3'-Dichlorobenzidine(DCB) has be shown carcinogenic in several animals, and the development of non-invasive biomonitoring method in workers exposed with it is a very important subject. DNA adduct is a good biomarker for biomonitoring about carcinogens exposure, and lymphocytes is a good non-invasive samples. So we studied to analyze metabolites in blood lymphocytes of female Sprague-Dawley rats exposed orally with DCB(20, 30, and 40 mg/kg wt.) for 3 weeks. For analysis of them, we isolated DNA adducts from blood lymphocytes by using the enzymes method in /sup 32/P-postlabeling, and measured them by using gas chromatography/mass spectrometry-selected ion monitoring(GC/MS-SIM). 4-aminobiphenyl and phenanthrene-d/sub 10/ were added as internal standard for blank sample. Standard metabolites of DCB were synthesized with using pyridine and acetic acid which were promoter and controller in acetylation of DCB. And they were used for calibration curve. Our results showed two kinds of metabolites in DNA adducts of blood lymphocytes. They were N-acetyl 3,3'-dichlorobenzidine(acDCB) and N,N'-diacetyl 3,3'-dichiorobenzidine(di-acDCB ). They were combined with DNA at the same time as an acetyl of it was removed. So we measured DCB and acDCB for two kinds of metabolites in DNA adducts of blood lymphocytes. Our results showed the levels of DCB were 1.46∼2.26 times more than that of acDCB. And also the levels of metabolites in 20, 30 and 40 mg/kg wt. were gradually increased with going days from 1st to 3rd week. They are 1.66, 1.38 and 0.90 times in total metabolites, 1.76, 1.49 and 1.02 times in DCB, and 1.51, 1.22 and 1.28 times in acDCB. In conclusion, the results of this study showed DCB exposed to rats formed DNA adduct in blood lymphocytes after acetylated to N-acetyl 3.3'-dichloro benzidine(acDCB) and N,N'-diacetyl 3,3'-dichlorobenzidine(di-acDCB), and they could be analyzed by us ing gas chromatography/mass spectrometry-selected ion monitoring(GC/MS-SIM).