• Title/Summary/Keyword: 제한효소분석

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Stock Identification of Todarodes pacificus in Northwest Pacific (북서태평양에 서식하는 살오징어(Todarodes pacificus) 계군 분석에 대한 고찰)

  • Kim, Jeong-Yun;Moon, Chang-Ho;Yoon, Moon-Geun;Kang, Chang-Keun;Kim, Kyung-Ryul;Na, Taehee;Choy, Eun Jung;Lee, Chung Il
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.17 no.4
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    • pp.292-302
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    • 2012
  • This paper reviews comparison analysis of current and latest application for stock identification methods of Todarodes pacificus, and the pros and cons of each method and consideration of how to compensate for each other. Todarodes pacificus which migrates wide areas in western North Pacific is important fishery resource ecologically and commercially. Todarodes pacificus is also considered as 'biological indicator' of ocean environmental changes. And changes in its short and long term catch and distribution area occur along with environmental changes. For example, while the catch of pollack, a cold water fish, has dramatically decreased until today after the climate regime shift in 1987/1988, the catch of Todarodes pacificus has been dramatically increased. Regarding the decrease in pollack catch, overfishing and climate changes were considered as the main causes, but there has been no definite reason until today. One of the reasons why there is no definite answer is related with no proper analysis about ecological and environmental aspects based on stock identification. Subpopulation is a group sharing the same gene pool through sexual reproduction process within limited boundaries having similar ecological characteristics. Each individual with same stock might be affected by different environment in temporal and spatial during the process of spawning, recruitment and then reproduction. Thereby, accurate stock analysis about the species can play an efficient alternative to comply with effective resource management and rapid changes. Four main stock analysis were applied to Todarodes pacificus: Morphologic Method, Ecological Method, Tagging Method, Genetic Method. Ecological method is studies for analysis of differences in spawning grounds by analysing the individual ecological change, distribution, migration status, parasitic state of parasite, kinds of parasite and parasite infection rate etc. Currently the method has been studying lively can identify the group in the similar environment. However It is difficult to know to identify the same genetic group in each other. Tagging Method is direct method. It can analyse cohort's migration, distribution and location of spawning, but it is very difficult to recapture tagged squids and hard to tag juveniles. Genetic method, which is for useful fishery resource stock analysis has provided the basic information regarding resource management study. Genetic method for stock analysis is determined according to markers' sensitivity and need to select high multiform of genetic markers. For stock identification, isozyme multiform has been used for genetic markers. Recently there is increase in use of makers with high range variability among DNA sequencing like mitochondria, microsatellite. Even the current morphologic method, tagging method and ecological method played important rolls through finding Todarodes pacificus' life cycle, migration route and changes in spawning grounds, it is still difficult to analyze the stock of Todarodes pacificus as those are distributed in difference seas. Lately, by taking advantages of each stock analysis method, more complicated method is being applied. If based on such analysis and genetic method for improvement are played, there will be much advance in management system for the resource fluctuation of Todarodes pacificus.

Analysis of Bone Marrow Micrometastasis Using RT-PCR in Patients with Small Cell Lung Carcinoma (소세포 폐암에서 역전사 중합효소 연쇄반응(RT-PCR)을 이용한 골수 미세전이(naicronaetastasis)의 분석)

  • Kim, Tae-You;Park, Jong-Kook;Ryoo, Baek-Ryeol;Im, Yung-Hyuck;Kang, Yoon-Koo
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.6
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    • pp.797-806
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    • 1999
  • Background: About 20% of small cell lung cancer(SCLC) patients have bone marrow(EM) metastasis at the time of diagnosis and the remaining patients are also considered with micrometastasis. In an attempt to detect EM micrometastasis, we used cytokeratin(CK)-20 as a molecular marker, which is specific for epithelial cells. Method: A sensitive RT-PCR assay was used to compare CK-20 expression both in SCLC cell line H209 and normal leukocyte and to evaluate EM aspirates of 28 SCLC patients. Result: H209 cell line showed CK-20 expression but normal leukocyte did not, suggesting CK-20 expression is lung tissue-specific. Of 28 patients(11 limited disease, 17 extensive disease), only 2(1/11, 1/17) samples tested revealed positive signal for CK-20. Two patients with CK-20 expression had EM metastasis or multiple bone involvement during follow-up. Conclusion: Although circulating tumor cells were detected in EM of small portion of patients with bone metastasis, CK-20 doesn't seem to be a reliable marker for the detection of micrometastasis in SCLC. This study emphasizes that identification of more specific marker for micrometastsis is mandatory prior to clinical application.

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Construction of the Genomic Expression Library of Bacillus anthracis for the Immunomic Analysis (면역체 분석을 위한 탄저균 유전자 발현 라이브러리의 구축)

  • Park, Moon-Kyoo;Jung, Kyoung-Hwa;Kim, Yeon-Hee;Rhie, Gi-Eun;Chai, Young-Gyu;Yoon, Jang-W.
    • Korean Journal of Microbiology
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    • v.46 no.1
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    • pp.21-26
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    • 2010
  • As the causative agent of Anthrax, Bacillus anthracis causes an acute fatal disease in herbivores such as cattle, sheep, and horses as well as humans. The therapeutics and prevention of anthrax currently available are based on antibiotics and the live attenuated vaccine strains, which may be problematic due to the emergency of antibiotic resistant strains or residual virulence in those vaccine strains. Therefore, it has been required to develop novel therapeutics and vaccines which are safer and applicable to humans. Recently, the development of the multivalent vaccine targeting both spores and vegetative cells of B. anthracis along with anthrax toxin has been reported. In our attempts to screen potential candidates for those multivalent vaccines, the whole genomic expression library of B. anthracis was constructed in this study. To the end, the partial digests of the genomic DNA from B. anthracis (ATCC 14578) with Sau3AI were ligated with the inducible pET30abc expression vectors, resulting in approximately $1{\times}10^5$ clones in E. coli BL21(DE3). The redundancy test by DNA nucleotide sequencing was performed for the randomly selected 111 clones and found 56 (50.5%) B. anthracis genes, 17 (15.3%) vector sequences, and 38 (34.2%) unknown genes with no sequence homology by BLAST. An inducible expression of the recombinant proteins was confirmed by Western blot. Interestingly, some clones could react with the antiserum against B. anthracis. These results imply that the whole genomic library constructed in this study can be applied for analyzing the immunomes of B. anthracis.

Human Impact on Sedimentary Environment of Estuarine Coastal Salt Marches, Southern Coastal Region of Korea Peninsula (인위적 환경변화에 따른 해안지역 퇴적환경의 변화)

  • 박의준
    • Journal of the Korean Geographical Society
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    • v.36 no.2
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    • pp.111-125
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    • 2001
  • An estuary is semi-inclosed inlets, located between terrestrial and marine environment. Since many estuaries along south-western coasts of Korean peninsula were affected by human settlements and activities, significant changes in sedimentation environments have been observed. The research area is divided into three distinct morpho-stratigraphic units: fluvial dominated area(Area1), mixed area(Area 2), tide-dominated area(Area3). The landform of this area has been changed by reclamation and river channel change. Temporal variations affected by dam construction, periodic freshet was iterrupted. Sediments began to continuously accmulate on estuary banks by tide. Meanwhile, because of the continuous but reduced discharge of fresh water, the salinity of estuarine sediments was declined. That processes made vegetated area( Phregmites lonivalvis and Suaeda japonica) to be expanded. It indicates that the magnitude and frequency of geomorphic processes has been significantly changed.

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A study of neuropeptides related to headaches in children with meningeal irritation signs (수막 자극 증상으로 내원한 환자에서 두통과 관련된 신경 펩타이드에 대한 연구)

  • Seo, Bo Gil;Yoo, Myung Hwan;Shim, Jae Won;Shim, Jung Yeon;Jung, Hye Lim;Park, Moon Soo;Kim, Deok-Soo
    • Clinical and Experimental Pediatrics
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    • v.49 no.5
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    • pp.533-538
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    • 2006
  • Purpose : The headache, one of the symptoms of meningitis, is related to abrupt elevation of intracranial pressure(ICP) or stimulation of intracranial nociceptive structure. However, in cases of mild elevation of ICP or normal findings of cerebrospinal fluid(CSF) analysis, patients sometimes complain of headaches. Therefore, other pathways may contribute to the occurrence of headaches in aseptic meningitis or meningismus. We intend to investigate the role of substance P(SP) and calcitonin gene-related peptide(CGRP) in aseptic meningitis or meningismus. Methods : We measured leukocyte count, the concentration of protein and glucose in CSF and ICP of patients with meningeal irritation sign. We also measured SP and CGRP levels by using immunoassay. We analyzed the relationship between the presence of headache and the value of SP and CGRP. Results : The concentrations of CGRP($18.8{\pm}10.5ng/mL$) in CSF and ICP($14.8{\pm}4.5cmH_2O$) in aseptic meningitis group were significantly higher than in those($14.1{\pm}7.4ng/mL$ and $12.0{\pm}5.1cmH_2O$, respectively) of the meningismus group(P<0.05). There was no significant difference in the SP levels between the two groups. In the aseptic meningitis group, the concentrations of SP and CGRP were significantly higher in the normal ICP group than in the elevated ICP group(P<0.05). Conclusion : Headaches in children with aseptic meningitis or meningismus is considered to be related to the elevation of the CSF levels of SP and CGRP.

Molecular Identification of Zoysia japonica and Zoysia sinica (Zoysia Species) Based on ITS Sequence Analyses and CAPS (ITS 염기서열 분석 및 CAPS를 이용한 조이시아 속(Zoysia) 들잔디와 갯잔디의 구별)

  • Hong, Min-Ji;Yang, Dae-Hwa;Jeong, Ok-Cheol;Kim, Yang-Ji;Park, Mi-Young;Kang, Hong-Gyu;Sun, Hyeon-Jin;Kwon, Yong-Ik;Park, Shin-Young;Yang, Paul;Song, Pill-Soon;Ko, Suk-Min;Lee, Hyo-Yeon
    • Horticultural Science & Technology
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    • v.35 no.3
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    • pp.344-360
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    • 2017
  • Zoysiagrasses are important turf plants used for school playgrounds, parks, golf courses, and sports fields. The two most popular zoysiagrass species are Zoysia japonica and Zoysia sinica. These are widely distributed across different growing zones and are morphologically distinguishable from each other; however, it is phenotypically difficult to differentiate those that grow along the coastal line from those in beach area habitats. A combination of morphological and molecular approaches is desirable to efficiently identify these two plant cultivars. In this study, we used a rapid identification system based on DNA barcoding of the nrDNA-internal transcribed spacer (ITS) regions. The nrDNA-ITS regions of ITS1, 5.8S nrDNA, and ITS2 from Z. japonica, Z. sinica, Agrostis stolonifera, and Poa pratensis were DNA barcoded to classify these grasses according to their molecular identities. The nrDNA-ITS sequences of these species were found at 686 bp, 687 bp, 683 bp, and 681 bp, respectively. The size of ITS1 ranged from 248 to 249 bp, while ITS2 ranged from 270 to 274 bp. The 5.8S coding region ranged from 163 - 164bp. Between Z. japonica and Z. sinica, nineteen (2.8%) nucleotide sites were variable, and the G+C content of the ITS region ranged from 55.4 to 63.3%. Substitutions and insert/deletion (indel) sites in the nrDNA-ITS sequence of Z. japonica and Z. sinica were converted to cleaved amplified polymorphic sequence (CAPS) markers, and applied to the Zoysia grasses sampled to verify the presence of these markers. Among the 62 control and collected grass samples, we classified three groups: 36 Z. japonica, 22 Z. sinica, and 4 Z. japonica/Z. sinica hybrids. Morphological classification revealed only two groups; Z. japonica and Z. sinica. Our results suggest that used of the nrDNA-ITS barcode region and CAPS markers can be used to distinguish between Z. japonica and Z. sinica at the species level.

8 Years Report of Urine Organic Acid Analysis - Comparison before and after Introduction of Neonatal Screening Test using Tandem Mass Spectrometry - (소변 유기산 분석 8년의 정리 -탠덤매스(Tandem mass spectrometry)를 이용한 신생아 선별검사 도입 전후의 비교-)

  • Ahn, Seok Min;Shin, Woo Chul;Jeong, Han Bin;Seo, Young Jun;Jeong, Hwal Rim;Yoon, Jong Hyung;Bae, Eun Ju;Lee, Hong Jin
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.18 no.1
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    • pp.1-12
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    • 2018
  • Purpose: Disorders of organic acid metabolism have various clinical manifestations and it may be life-threatening. The prognoses of affected children are dependent on early diagnosis and treatment. We report this study to find out detection rate of referred samples, clinical manifestations and age distribution after introduction of neonatal screening test using tandem mass spectrometry in Hallym University Chuncheon Sacred Heart Hospital during 8 years and 9 months. Methods: The 2,794 patients referred from Jan. 2007 to Sep. 2015 were divided into four groups according to age. We conducted organic acid analysis of urine samples of patients and analyzed clinical manifestations and distributions of age at the diagnosis. For patients with ambiguous results, reanalysis of urine organic acid after diet restriction, protein loading and restriction, has been done. Results: A total of 626 patients with 20 disorders were diagnosed. Mitochondrial disorders (482 patients) were the most common diagnosis, followed by ketolytic defects (67), 3-hydroxyisobutyric aciduria (32), EPEMA syndrome (8), 3-methylcrotonyl glycinuria (7), glutaric aciduria type II (6) and type I (4), methylmalonic aciduria (3), isovaleric aciduria (3) and medium chain acyl-CoA dehydrogenase deficiency (3). Conclusion: As neonatal screening test using tandem mass spectrometry is increasingly common and medical environment is changed, detection rate of disorders of organic acid metabolism in this study has decreased compared to previous report. Because the deterioration can be prevented by early diagnosis and treatment, many pediatricians have to pay special attention to possibility of the disorders and make an effort for early diagnosis in clinical setting.

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Identification and Characterization of Wild Yeasts Isolated from Korean Domestic Grape Varieties (국산 포도로부터 분리한 야생효모의 동정 및 특성)

  • Choi, Sang-Hoon;Hong, Young-Ah;Choi, Yoon-Jung;Park, Heui-Dong
    • Food Science and Preservation
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    • v.18 no.4
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    • pp.604-611
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    • 2011
  • Several wild yeasts were isolated from Korean grape varieties before and during spontaneous fermentation. Among them, four strains were isolated based on the alcohol content and flavor production in wine after fermentation of apple juice. In this study, the four yeast strains were identified and characterized. PCR-restriction fragment length polymorphism analysis of ITS I-5.8S-ITS II region with restriction endonuclease Hae III and Hinf I resulted in that all the strains showed a typical pattern of Saccharomyces cerevisiae. Pulse field gel electrophoresis showed three different chromosome patterns with a same band between strains SS89 and SS812. When ITS I-5.8S-ITS II sequences of the four strains were compared with one another, they were similar to those of Saccharomyces cerevisiae CBS 4054 type strain. Identity of the sequences was higher than 97% with those of the type strain. Phylogenetic analysis showed based on the sequences showed they were genetically closed to the type strain. The four identified strains were tested in a medium containing 200 ppm potassium metabisulfite, and the MM10 and WW108 inhibition rates resulted at up to 24 h. The four strains were tested at an incubation temperature of $30^{\circ}C$. The 30% sugar concentration in the medium (w/v) showed the highest growth in 36 h, especially in the case of SS89, which was close to growth 40. The four strains were tested in an 8% ethanol medium (v/v). Alcohol tolerance was initially kept in the incubation process. The strains began to adapt, however, to the exceeded resistance. The four strains showed the lowest inhibition rate at 24 h.

Chronic Hereditary Tyrosinemia Type I with Novel Mutation in FAH Gene (FAH gene novel mutation을 가진 만성형 Hereditary tyrosinemia 1형)

  • Yang, Sungmin;Choi, Hyo Won;Kang, Yun Koo;Lee, Jin-Sung;Namgoong, Mee Kyung
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.20 no.2
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    • pp.55-62
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    • 2020
  • A 22-month-old girl who had taken iron supplements due to iron deficiency anemia, presented bloody mucoid stool for one month. She had a bruise at the right periorbital area due to minor trauma and hepatosplenomegaly. Laboratory studies showed anemia, thrombocytopenia, elevated alkaline phosphatase (ALP), hypophosphatemia, decreased haptoglobin, hypocomplementemia, negative direct/indirect Coomb's test, normal vitamin D3 level and high PTHi. Wrist x-ray showed no signs of rickets. The abdominal ultrasound showed only accessory spleen. Tandem mass spectrometry was normal. During follow up, bloody stool regressed after seven days of withdrawal of iron supplement and cow milk, and the total CO2 level had been within 15-20 mEq/L with normal anion gap. NGS (next generation sequencing) panel test for evaluation of renal tubular acidosis showed negative results. After low dose steroid and vitamin D supplements under the impression of hypocomplementemic vasculitis, thrombocytopenia, C3/C4, decreased haptoglobin, and elevated ALP level became normal. At 57 months of age, laboratory findings showed elevated liver enzyme, ALP and gamma-glutamyl transferase again. And liver cirrhosis with splenomegaly and diffuse renal disease were reported with abdomen CT scan. Liver biopsy reported macro- and micronodular cirrhosis. Urine organic acid profile showed elevated succinylacetone level. Whole exome sequencing revealed novel compound heterozygous mutations (NM_00137.2:c.107T>C, NM_00137, 2:c.614T>C) in FAH gene and confirmed by Sanger sequencing. Consequently, the patient was diagnosed as chronic hereditary tyrosinemia type I. She started low phenylalanine/tyrosine diet and nitisinone treatment. Our case had presented symptoms very slowly, which is the first case of chronic tyrosinemia type I in South Korea.

Construction of Genetic Linkage Map and Identification of Quantitative Trait Loci in Populus davidiana using Genotyping-by-sequencing (Genotyping-by-sequencing 기법을 이용한 사시나무(Populus davidiana) 유전연관지도 작성 및 양적형질 유전자좌 탐색)

  • Suvi Kim;Yang-gil Kim;Dayoung Lee;Hye-jin Lee;Kyu-Suk Kang
    • Journal of Korean Society of Forest Science
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    • v.112 no.1
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    • pp.40-56
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    • 2023
  • Tree species within the Populus genus grow rapidly and have an excellent capacity to absorb carbon, conferring substantial ability to effective purify the environment. Poplar breeding can be achieved rapidly and efficiently if a genetic linkage map is constructed and quantitative trait loci (QTLs) are identified. Here, a high-density genetic linkage map was constructed for the control pollinated progeny using the genotyping-by-sequencing (GBS) technique, which is a next-generation sequencing method. A search was also performed for the genes associated with quantitative traits located in the genetic linkage map by examining the variables of height and diameter at root collar, and resilience to insect damage. The height and diameter at root collar were measured directly, while the ability to recover from insect damage was scored in a 4-year-old breeding population of aspen hybrids (Odae19 × Bonghyeon4 F1) established in the research forest of Seoul National University. After DNA extraction, paternity was confirmed using five microsatellite markers, and only the individuals for which paternity was confirmed were used for the analysis. The DNA was cut using restriction enzymes and the obtained DNA fragments were prepared using a GBS library and sequenced. The analyzed results were sorted using Populus trichocarpa as a reference genome. Overall, 58,040 aligned single-nucleotide polymorphism (SNP) markers were identified, 17,755 of which were used for mapping genetic linkages. The genetic linkage map was divided into 19 linkage groups, with a total length of 2,129.54 cM. The analysis failed to identify any growth-related QTLs, but a gene assumed to be related to recovery from insect damage was identified on linkage group (chromosome) 4 through genome-wide association study.