• Title/Summary/Keyword: fingerprinting

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Identification of Cold Stress-related Proteins in Rice Leaf Tissue (벼의 잎 조직에서 발현되는 저온 스트레스 관련 단백질의 분리 동정)

  • Lee Dong-Gi;Lee Sang-Hoon;Lee Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.25 no.4
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    • pp.287-296
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    • 2005
  • To investigation protein expression pattern in rice leaves exposed to cold stress, the soluble proteins extracted from leaf tissue were fractionated with $15\%$ PEG and separated by two-dimensional polyacrylamide gel electrophoresis (2-DE). Differentially expressed proteins were identified by peptide mass fingerprinting using matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Eight proteins up-regulated and 10 down-regulated were found in $15\%$ PEG supernatant fraction. In addition, 13 proteins up-regulated and 14 down-regulated were found in $15\%$ PEG pellet fraction. It was identified the differentially expressed proteins in $15\%$ PEG supernatant fraction as pimerase/dehydratase fructokinase, ribose-5-phosphate isomerase (Rpi), chaperonin 21 precursor, probable photosystem II oxygen-envolving complex (PS II OEC) protein 2 precursor and thioredoxin h-type (Trx-h) and those in $15\%$ PEG pellet fraction as OSINBb0059K02.15, hypothetical protein, putative mitogen-activated protein kinase kinase (MAPKK), beta 7 subunit of 205 proteasome, ribulose-1, 5-bisphosphate carboxylase/oxygenase (Rubisco) small subunit. These proteins are involved in metabolism, energy, protein synthesis, disease/defense and signal transduction-related proteins.

Identification of Korean Native Goat Meat using Amplified Fragment Length Polymorphism (AFLP) DNA Markers (Amplified Fragment Length Polymorphism (AFLP) DNA Marker를 이용한 한국 재래흑염소육 감별)

  • 정의룡
    • Food Science of Animal Resources
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    • v.22 no.4
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    • pp.301-309
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    • 2002
  • This study was carried out to develop the breed-specific DNA markers for breed identification of Korean native goat meat using amplified fragment length polymorphism (AFLP)-PCR techniques. The genomic DNAs of Korean native goat, imported black goat and four dairy goat breeds(Saanen, Alpine, Nubian and Toggenburg) were extracted from muscle tissues or blood. Genomic DNA was digested with a particular combination of two restriction enzymes with 4 base(Mse I and Taq I) and 6 base(EcoR I and Hind III) recognition sites, ligated to restriction specific adapters and amplified using the selective primer combinations. In AFLP profiles of polyacrylamide gels, the number of scorable bands produced per primer combination varied from 36 to 74, with an average of 55.5. A total of 555 bands were produced, 149(26.8%) bands of which were polymorphic. Among the ten primer combinations, two bands with 2.01 and 1.26 kb in M13/H13 primer and one band with 1.65 kb in E35/H14 primer were found to be breed-specific AFLP markers in Korean native goat when DNA bands were compared among the goat breeds. In the E35/H14 primer combination, 2.19, 2.03, 0.96 and 0.87 kb bands detected in imported black goat, 2.13 kb band in Saanen breed and 2.08 kb band in Nubian breed were observed as breed-specific bands showing differences between goat breeds, respectively. The E35/H14 primer combination produced four DNA bands distinguished between Korean native goat and Saanen breed. The is study suggested that the breed specific AFLP bands could be used as DNA markers for the identification of Korean native goat meat from imported black goat and dairy goat meats.

Restriction Fragment Length Polymorphism of Mycobacterium Tuberculosis in a University Hospital in Seoul (서울의 한 대학병원에서 동정된 결핵균 균주의 RFLP 양상)

  • Kim, Woo-Jin;Yim, Jae-Joon;Lee, Jae-Ho;Yoo, Chul-Gyu;Chung, Hee-Soon;Han, Sung-Koo;Shim, Young-Soo;Kim, Young-Whan
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.3
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    • pp.308-314
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    • 2000
  • Background : DNA fingerprinting of Mycobacterium tuberculosis with RFLP is a very useful tool for deciphering the molecular epidemiology of tuberculosis. An international comparison of the RFLP pattern became possible with the proposal to standardize RFLP methodology using Pvu-II restricted IS-6110, and the comparison has noted some predominance of RFLP pattern in East Asia. The RFLP patterns of tuberculosis strains collected at SNUH was studied and was compared with other strains from East Asia. Method : Fifty strains of M. tuberculosis were isolated from patients who visited 'or were admitted to the SNUH in 1998. Some isolates belonging to the Beijing family were also received. After the extraction of DNA from M. tuberculosis isolates, the chromosomal DNAs were digested with Pvu-II and analyzed by the Southern blot method with DNA probe to IS6110. Result : Six strains belonged to the Beijing family. The RFLP patterns of other 9 strains were similar to each other. No statistically significant endobronchial tuberculosis, presence of underlying disease. and the province of residence were found. Conclusion : Few groups of M. tuberculosis strains from SNUH showed similar RFLP patterns, but their clinical implications are not yet clear.

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Biphasic Study to Characterize Agricultural Biogas Plants by High-Throughput 16S rRNA Gene Amplicon Sequencing and Microscopic Analysis

  • Maus, Irena;Kim, Yong Sung;Wibberg, Daniel;Stolze, Yvonne;Off, Sandra;Antonczyk, Sebastian;Puhler, Alfred;Scherer, Paul;Schluter, Andreas
    • Journal of Microbiology and Biotechnology
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    • v.27 no.2
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    • pp.321-334
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    • 2017
  • Process surveillance within agricultural biogas plants (BGPs) was concurrently studied by high-throughput 16S rRNA gene amplicon sequencing and an optimized quantitative microscopic fingerprinting (QMF) technique. In contrast to 16S rRNA gene amplicons, digitalized microscopy is a rapid and cost-effective method that facilitates enumeration and morphological differentiation of the most significant groups of methanogens regarding their shape and characteristic autofluorescent factor 420. Moreover, the fluorescence signal mirrors cell vitality. In this study, four different BGPs were investigated. The results indicated stable process performance in the mesophilic BGPs and in the thermophilic reactor. Bacterial subcommunity characterization revealed significant differences between the four BGPs. Most remarkably, the genera Defluviitoga and Halocella dominated the thermophilic bacterial subcommunity, whereas members of another taxon, Syntrophaceticus, were found to be abundant in the mesophilic BGP. The domain Archaea was dominated by the genus Methanoculleus in all four BGPs, followed by Methanosaeta in BGP1 and BGP3. In contrast, Methanothermobacter members were highly abundant in the thermophilic BGP4. Furthermore, a high consistency between the sequencing approach and the QMF method was shown, especially for the thermophilic BGP. The differences elucidated that using this biphasic approach for mesophilic BGPs provided novel insights regarding disaggregated single cells of Methanosarcina and Methanosaeta species. Both dominated the archaeal subcommunity and replaced coccoid Methanoculleus members belonging to the same group of Methanomicrobiales that have been frequently observed in similar BGPs. This work demonstrates that combining QMF and 16S rRNA gene amplicon sequencing is a complementary strategy to describe archaeal community structures within biogas processes.

FingerPrint building method using Splite-tree based on Indoor Environment (실내 환경에서 WLAN 기반의 Splite-tree를 이용한 가상의 핑거 프린트 구축 기법)

  • Shin, Soong-Sun;Kim, Gyoung-Bae;Bae, Hae-Young
    • Journal of the Korea Society of Computer and Information
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    • v.17 no.6
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    • pp.173-182
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    • 2012
  • A recent advance in smart phones is increasing utilization of location information. Existing positioning system was using GPS location for positioning. However, the GPS cannot be used indoors, if GPS location has an incorrectly problem. In order to solve indoor positioning problems of indoor location-based positioning techniques have been investigated. There are a variety of techniques based on indoor positioning techniques like as RFID, UWB, WLAN, etc. But WLAN location positioning techniques take advantage the bond in real life. WLAN indoor positioning techniques have a two kind of method that is centroid and fingerprint method. Among them, the fingerprint technique is commonly used because of the high accuracy. In order to use fingerprinting techniques make a WLAN signal map building that is need to lot of resource. In this paper, we try to solve this problem in an Indoor environment for WLAN-based fingerprint of a virtual building technique, which is proposed. Proposed technique is classified Cell environment in existed Indoor environment, all of fingerprint points are shown virtual grid map in each Cell. Its method can make fingerprint grid map very quickly using estimate virtual signal value. Also built signal value can take different value depending of the real estimate value. To solve this problem using a calibration technique for the Splite-tree is proposed. Through calibration technique that improves the accuracy for short period of time. It also is improved overall accuracy using predicted value of around position in cell.

Improvement of PCR Amplification Bias for Community Structure Analysis of Soil Bacteria by Denaturing Gradient Gel Electrophoresis

  • Ahn, Jae-Hyung;Kim, Min-Cheol;Shin, Hye-Chul;Choi, Min-Kyeong;Yoon, Sang-Seek;Kim, Tae-Sung;Song, Hong-Gyu;Lee, Geon-Hyoung;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1561-1569
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    • 2006
  • Denaturing gradient gel electrophoresis (DGGE) is one of the most frequently used methods for analysis of soil microbial community structure. Unbiased PCR amplification of target DNA templates is crucial for efficient detection of multiple microbial populations mixed in soil. In this study, DGGE profiles were compared using different pairs of primers targeting different hypervariable regions of thirteen representative soil bacteria and clones. The primer set (1070f-1392r) for the E. coli numbering 1,071-1,391 region could not resolve all the 16S rDNA fragments of the representative bacteria and clones, and moreover, yielded spurious bands in DGGE profiles. For the E. coli numbering 353-514 region, various forward primers were designed to investigate the efficiency of PCR amplification. A degenerate forward primer (F357IW) often yielded multiple bands for a certain single 16S rDNA fragment in DGGE analysis, whereas nondegenerate primers (338f, F338T2, F338I2) differentially amplified each of the fragments in the mixture according to the position and the number of primer-template mismatches. A forward primer (F352T) designed to have one internal mismatch commonly with all the thirteen 16S rDNA fragments efficiently produced and separated all the target DNA bands with similar intensities in the DGGE profiles. This primer set F352T-519r consistently yielded the best DGGE banding profiles when tested with various soil samples. Touchdown PCR intensified the uneven amplification, and lowering the annealing temperature had no significant effect on the DGGE profiles. These results showed that PCR amplification bias could be much improved by properly designing primers for use in fingerprinting soil bacterial communities with the DGGE technique.

Proteomic Analyses of Chinese Cabbage(Brassica campestris L. pekinensis) Affected by High Temperature Stresses in Highland Cultivation During Summer in Korea (Proteomics를 이용한 고랭지 배추의 고온장해 해석)

  • Shin, Pyung-Gyun;Hong, Sung-Chang;Chang, An-Cheol;Kim, Sang-Hyo;Lee, Ki-Sang
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1649-1653
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    • 2007
  • High temperature stresses have caused growth inhibition and delayed heading in highland cultivation Chinese cabbage during summer in Korea. We have studied high temperature stress responses in the terms of changes of inorganic components and proteins by proteomic analyses. Insufficiencies of nitrogen and phosphorus have affected growth rate and calcium deficiency has caused blunted heading. Proteins extracted from Brassica seedling grown at the altitude of 600m and 900m in the Mount Jilun were extracted and analysed by 2-dimentional polyacrylamide gel electrophoresis. Profiles of protein expression was then analyzed by 2-dimentional gel analyses. Protein spots showing different expression level were picked using the spot handling workstation and subjected to MALDI-TOF MS. Total 48 protein spots were analyzed by MALDI-TOF MS and 30 proteins spots out of 48 were identified by peptide mass fingerprinting analyses. Fourteen proteins were up-regulated in extracts from the altitude of 900m and they were identified as oxygen-evolving proteins, rubisco activase and ATPase etc. Sixteen proteins were up-regulated in extracts from the altitude of 600m and they were identified as glutathione S-transferase(1, 28kD cold induced- and 24 kD auxin-binding proteins) and salt-stress induced protein etc. These stress-induced proteins were related to the mediated protective mechanism against oxidative damage during various stresses. The results indicated that physiological phenomenon in response to high temperature stresses might be resulted by complex and multiple array of responses with drought, heat, oxidative, salt, and cold by high temperature.

Characteristics of Dioscorea alata L. Introduced from Tropical and Subtropical Regions (도입 마(Dioscorea alata L.)의 특성 분석)

  • Chang, K.J.;Yoo, K.O.;Park, C.H.;Park, J.I.;Hong, K.H.;Park, J.H.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.3 no.1
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    • pp.48-69
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    • 2001
  • A lot of clones of the genus Dioscorea have been introduced from some tropical and subtropical regions since 1997. In 33 clones of water yams (Dioscorea alata L.), some morphological characteristics were investigated at the field. Variation ranges of the total weight and tuber number per stump were within the ranges from 90 to 2,147 g with an average of 610 g ; and 1.3-4.7 with an average of 2.8, respectively. The color tones observed on the tuber-flesh were sorted into 3 color-categories, i.e., white, pale brown and pale purple, and those on leaves were sorted into 3 color-categories, i.e., green, heavy green and purplish green. Intraspecific genetic relationship of 19 variation types of the Yam classified by their external morphological characteristics such as leaf and tuber shape was assessed by DNA using random and specific primers. Twenty two out of 113 primers (100 random[10-mer] primers, two 15 mer [M13 core sequence, and (GGAT)4 sequence]) had been used in PCR-amplification. Only 12 primers, however, were successful in DNA amplification in all of the analyzed plants, resulting in 93 randomly and specifically amplified DNA fragments. The analyzed taxa showed very high polymorphisms(69 bands, 71.0%), allowing individual taxon to be identified based on DNA fingerprinting. Monomorphic bands among total amplified DNA bands of each primer was low under the 50%. Similarity indices between accessions were computed from PCR(polymerase chain reaction) data, and genetic relationships among intraspecific variations were closely related at the levels ranging from 0.66 to 0.90.

Application of the Latest European Standard(EN 15522-2) for Marine Oil Spill Analysis: A Study on its Effectiveness in Analyzing Samples from Korean Incidents (해양 기름유출사고 분석을 위한 최신 유럽표준(EN 15522-2) 적용: 한국 사고 샘플 분석의 효율성에 관한 연구)

  • Youjeong Park;Duwon Lee;Heejin Lee
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.30 no.1
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    • pp.58-64
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    • 2024
  • Approximately 250 marine oil spill accidents have occurred in Korea, with profound impact on local communities and the environment. The restoration process necessitates significant resources and costs to return affected areas to their pre-accident state. In accordance with the polluter pays principle, compensation is demanded from polluter, as stipulated in both international conventions and national laws. Consequently, investigations are conducted to determine civil and criminal liability. As the importance of investigation actors in oil spill accidents increases, standards such as CEN 15522-2 and ASTM D 3248 are employed to determine the similarity between the spilled oil and the oil of the suspected ship. Among these standards, CEN 15522-2, the most actively used European standard, underwent its third revision and is now known as EN 15522-2, as of 2023. This study used EN 15522-2 to analyze samples from marine oil spill accidents that occurred in Korea. The results indicated that, considering the characteristics of domestic spills where light fuel oil spills account for more than 40%, the application of EN 15522-2, which includes low-boiling point substances such as Adamantanes, was confirmed to be highly effective.

Discrimination of African Yams Containing High Functional Compounds Using FT-IR Fingerprinting Combined by Multivariate Analysis and Quantitative Prediction of Functional Compounds by PLS Regression Modeling (FT-IR 스펙트럼 데이터의 다변량 통계분석을 이용한 고기능성 아프리칸 얌 식별 및 기능성 성분 함량 예측 모델링)

  • Song, Seung Yeob;Jie, Eun Yee;Ahn, Myung Suk;Kim, Dong Jin;Kim, In Jung;Kim, Suk Weon
    • Horticultural Science & Technology
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    • v.32 no.1
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    • pp.105-114
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    • 2014
  • We established a high throughput screening system of African yam tuber lines which contain high contents of total carotenoids, flavonoids, and phenolic compounds using ultraviolet-visible (UV-VIS) spectroscopy and Fourier transform infrared (FT-IR) spectroscopy in combination with multivariate analysis. The total carotenoids contents from 62 African yam tubers varied from 0.01 to $0.91{\mu}g{\cdot}g^{-1}$ dry weight (wt). The total flavonoids and phenolic compounds also varied from 12.9 to $229{\mu}g{\cdot}g^{-1}$ and from 0.29 to $5.2mg{\cdot}g^{-1}$dry wt. FT-IR spectra confirmed typical spectral differences between the frequency regions of 1,700-1,500, 1,500-1,300 and $1,100-950cm^{-1}$, respectively. These spectral regions were reflecting the quantitative and qualitative variations of amide I, II from amino acids and proteins ($1,700-1,500cm^{-1}$), phosphodiester groups from nucleic acid and phospholipid ($1,500-1,300cm^{-1}$) and carbohydrate compounds ($1,100-950cm^{-1}$). Principal component analysis (PCA) and subsequent partial least square-discriminant analysis (PLS-DA) were able to discriminate the 62 African yam tuber lines into three separate clusters corresponding to their taxonomic relationship. The quantitative prediction modeling of total carotenoids, flavonoids, and phenolic compounds from African yam tuber lines were established using partial least square regression algorithm from FT-IR spectra. The regression coefficients ($R^2$) between predicted values and estimated values of total carotenoids, flavonoids and phenolic compounds were 0.83, 0.86, and 0.72, respectively. These results showed that quantitative predictions of total carotenoids, flavonoids, and phenolic compounds were possible from FT-IR spectra of African yam tuber lines with higher accuracy. Therefore we suggested that quantitative prediction system established in this study could be applied as a rapid selection tool for high yielding African yam lines.