• Title/Summary/Keyword: Fingerprinting analysis

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Proteomic Analysis of the Hydrophobic Fraction of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood

  • Jeong, Ju Ah;Lee, Yoon;Lee, Woobok;Jung, Sangwon;Lee, Dong-Seong;Jeong, Namcheol;Lee, Hyun Soo;Bae, Yongsoo;Jeon, Choon-Ju;Kim, Hoeon
    • Molecules and Cells
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    • v.22 no.1
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    • pp.36-43
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    • 2006
  • Mesenchymal stem cells (MSCs) are promising candidates for cell therapy and tissue engineering, but their application has been impeded by lack of knowledge of their core biological properties. In order to identify MSC-specific proteins, the hydrophobic protein fraction was individually prepared from two different umbilical cord blood (UCB)-derived MSC populations; these were then subjected to two-dimensional (2D) gel electrophoresis and peptide mass fingerprinting matrix-assisted laser desorption/ionization (MALDI)-time of flight (TOF)-mass spectrometry (MS). Although the 2D gel patterns differed somewhat between the two samples, computer-assisted image analysis identified shared protein spots. 35 spots were reliably identified corresponding to 32 different proteins, many of which were chaperones. Based on their primary sub-cellular locations the proteins could be grouped into 6 categories: extracellular, cell surface, endoplasmic reticular, mitochondrial, cytoplasmic and cytoskeletal proteins. This map of the water-insoluble proteome may provide valuable insights into the biology of the cell surface and other compartments of human MSCs.

Phylogenetic Analysis of Bacterial Diversity in the Marine Sponge, Asteropus simplex, Collected from Jeju Island (제주도에서 채집한 해양 해면, Asteropus simplex의 공생세균에 관한 계통학적 분석)

  • Jeong, In-Hye;Park, Jin-Sook
    • Korean Journal of Microbiology
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    • v.48 no.4
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    • pp.275-283
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    • 2012
  • Culture-dependent RFLP and culture-independent DGGE were employed to investigate the bacterial community associated with the marine sponge Asteropus simplex collected from Jeju Island. A total of 120 bacterial strains associated with the sponge were cultivated using modified Zobell and MA media. PCR amplicons of the 16S rDNA from the bacterial strains were digested with the restriction enzymes HaeIII and MspI, and then assigned into different groups according to their restriction patterns. The 16S rDNA sequences derived from RFLP patterns showed more than 94% similarities compared with known bacterial species, and the isolates belonged to five phyla, Alphaproteobacteria, Gammaproteobacteria Actinobacteria, Bacteroidetes, and Firmicutes, of which Gammaproteobacteria was dominant. DGGE fingerprinting of 16S rDNAs amplified from the sponge-derived total gDNA showed 12 DGGE bands, and their sequences showed more than 90% similarities compared with available sequences. The sequences derived from DGGE bands revealed high similarity with the uncultured bacterial clones. DGGE revealed that bacterial community consisted of seven phyla, including Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Deltaproteobacteria, Actinobacteira, Chloroflexi, and Nitrospira. Alphaproteobacteria, Gammaproteobacteria, and Actinobacteria were commonly found in bacteria associated with A. simplex by both RFLP and DGGE methods, however, overall bacterial community in the sponge differed depending on the analysis methods. Sponge showed more various bacterial community structures in culture-independent method than in culture-dependent method.

Use of Microsatellite Markers Derived from Genomic and Expressed Sequence Tag (EST) Data to Identify Commercial Watermelon Cultivars (수박 시판 품종의 식별을 위한 Genomic과 Expressed Sequence Tag (EST)에서 유래된 Microsatellite Marker의 이용)

  • Kwon, Yong-Sham;Hong, Jee-Hwa;Kim, Du-Hyun;Kim, Do-Hoon
    • Horticultural Science & Technology
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    • v.33 no.5
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    • pp.737-750
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    • 2015
  • This study was carried out to construct a DNA profile database for 102 watermelon cultivars through the comparison of polymorphism level and genetic relatedness using genomic microsatellite (gMS) and expressed sequence tag (EST)-microsatellite (eMS) markers. Sixteen gMS and 10 eMS primers showed hyper-variability and were able to represent the genetic variation within 102 watermelon cultivars. With gMS markers, an average of 3.63 alleles per marker were detected with a polymorphism information content (PIC) value of 0.479, whereas with eMS markers, the average number of alleles per marker was 2.50 and the PIC value was 0.425, indicating that eMS detects a lower polymorphism level compared to gMS. Cluster analysis and Jaccard's genetic distance coefficients using the unweighted pair group method with arithmetic average (UPGMA) based on the gMS, eMS, and combined data sets showed that 102 commercial watermelon cultivars could be categorized into 6 to 8 major groups corresponding to phenotypic traits. Moreover, this method was sufficient to identify 78 out of 102 cultivars. Correlation analysis with Mantel tests for those clusters using 3 data sets showed high correlation ($r{\geq}0.80$). Therefore, the microsatellite markers used in this study may serve as a useful tool for germplasm evaluation, genetic purity assessment, and fingerprinting of watermelon cultivars.

Standardization and HPTLC Fingerprinting of a Polyherbal Unani Formulation

  • Beg, Mirza Belal;Viquar, Uzma;Naikodi, Mohammad Abdul Rasheed;Suhail, Habiba;Kazmi, Munawwar Husain
    • CELLMED
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    • v.11 no.1
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    • pp.4.1-4.8
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    • 2021
  • Background: The Unani system of medicine has been practised since centuries for the treatment of a range of diseases. In spite of their efficacy they have been widely criticised due to the lack of standardization and poor quality control. Standardization of Unani medicine is a valuable issue at the present because they are very prone to contamination, deterioration, adulteration and variation in composition due to biodiversity as well as careless collection. Objective: To Standardize and Development of HPTLC Fingerprinting of a polyherbal Unani formulation Qurs-e-Safa. Materials and methods: The conventional and modern analytical techniques were used to standardise Qurs-e-Safa. The study was carried into three different batches of Qurs-e-Safa prepared with its ingredients. The parameters studied are organoleptic, microscopic, physicochemical parameters, phytochemical screening, TLC, HPTLC profile, aflatoxin, microbial load and heavy metal analysis. Results and conclusion: Qurṣ-e-Sa'fa is dark yellow in colour and aromatic smell. Uniformity of diameter and weight variation were found to be 13 ± 0, and 524.7 ± 1.72 mg. friability, hardness and disintegration time of all 3 batches were found to be (0.0615 ± 0.004, 0.0885 ± 0.0047 and 0.0725 ± 0.0058), (3.5 ± 0.2886, 3.67 ± 0.1674 and 3.67 ± 0.1674) and (16 to 17 minutes). Extractive value were found to be maximum in distilled water (38.488 ± 0.20, 37.3824 ± 0.38 and 39.8177 ± 0.13) followed by alcohol (27.5406 ± 0.54, 27.5656 ± 0.32 and 26.9229 ± 0.25). Loss of weight on drying, pH, total ash, acid insoluble ash, qualitative test was set in. Phytochemical screening revealed the presence of Carbohydrates, Phenols, Resins, Proteins, Steroids, fixed oil and Flavonoids. The microbial load was found absent and heavy metals were within permissible limits. The data evolved from the study may serve as a reference to validate and also help in the quality control of other finished products in future research.

The genetic structure of taro: a comparison of RAPD and isozyme markers

  • Sharma, Kamal;Mishra, Ajay Kumar;Misra, Raj Shekhar
    • Plant Biotechnology Reports
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    • v.2 no.3
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    • pp.191-198
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    • 2008
  • Germplasm characterization and evolutionary process in viable populations are important links between the conservation and utilization of plant genetic resources. Here, an investigation is made, based on molecular and biochemical techniques for assessing and exploiting the genetic variability in germplasm characterization of taro, which would be useful in plant breeding and ex situ conservation of taro plant genetic resources. Geographical differentiation and phylogenetic relationships of Indian taro, Colocasia esculenta (L.) Schott, were analyzed by random amplified polymorphic DNA (RAPD) and isozyme of seven enzyme systems with specific reference to the Muktakeshi accession, which has been to be proved resistant to taro leaf blight caused by P. colocasiae. The significant differentiations in Indian taro cultivars were clearly demonstrated by RAPD and isozyme analysis. RAPD markers showed higher values for genetic differentiation among taro cultivars and lower coefficient of variation than those obtained from isozymes. Genetic differentiation was evident in the taro accessions collected from different regions of India. It appears that when taro cultivation was introduced to a new area, only a small fraction of genetic variability in heterogeneous taro populations was transferred, possibly causing random differentiation among locally adapted taro populations. The selected primers will be useful for future genetic analysis and provide taro breeders with a genetic basis for selection of parents for crop improvement. Polymorphic markers identified in the DNA fingerprinting study will be useful for screening a segregating population, which is being generated in our laboratory aimed at developing a taro genetic linkage map.

Development of Reactor Vessel Head Penetration Performance Demonstration System in Korea (국내 원자로 상부헤드관통관 기량검증 기술개발)

  • Kim, Yongsik;Yoon, Byungsik;Yang, Seunghan
    • Transactions of the Korean Society of Pressure Vessels and Piping
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    • v.10 no.1
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    • pp.44-50
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    • 2014
  • There were many flaw issues of reactor vessel head penetration in USA fleets. USNRC issued 10CFR50.55a to implement reactor vessel head penetration ultrasonic examination performance demonstration(PD) in US for enhancement of inspection reliability. After September 2009, all US utilities inspected their RVHP with PD qualified system. Korea Hydro and Nuclear Power Company(KHNP) have developed reactor vessel head penetration performance demonstration system for ultrasonic test to apply for pressurized light-water reactor power plants in accordance with 10CFR50.55a since September 2011. RVHP configuration surveying and analysis, code requirement analysis, and performance demonstration specimen design were performed up to this day. Fingerprinting of manufactured specimen, development of test data management program, development of operation procedure, input of flawed data, and development of final report will be performed for the next step. This paper describes the development status of the performance demonstration system for reactor vessel head penetration ultrasonic examination in Korea.

Molecular fingerprinting of olive flounder pathogenic Streptococcus parauberis strains by random amplified polymorphic DNA analysis

  • Jung, Yong-Uk;Kang, Sang-Hyuck;Jin, Chang-Nam;Kang, Bong-Jo;Heo, Moon-Soo
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.899-903
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    • 2005
  • Two infectious species of Streptococcosis pathogens were detected by multiplex PCR assay. Detection rates of Streptococcus iniae and S. parauberis could reach 44.9% and 55.1% respectively for one year during 2004 to 2005 in Jeju island. These findings showed that S. parauberis strains were important pathogen with streptococcosis of olive flounder in Jeju island. These findings showed that S. parauberis strains were important pathogen with streptococcosis of olive flounder in Jeiu island. In the present study we have investigated the interspecific relationship of all Jeju area of S. parauberis by RAPD analysis. Represent strains divided to four groups by RAPD fingerprints. The important differences observed between the olive flounder isolates suggest that they could constitute a well-differentiated group or a separate clonal line within this bacterial species. Though, serological research of S. parauberis strains in Jeju island not exist yet. These strains doing the serological evolution.

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Chemotype Discrimination and Rapid Identification of Angelica Roots by DART-TOF-MS

  • Kim, Hye-Jin;Piao, Xiang-Lan;Jang, Young-Pyo
    • Natural Product Sciences
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    • v.17 no.3
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    • pp.202-205
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    • 2011
  • The Angelica root has been used as a medicinal herb in many Asian countries including Korea, China, and Japan. Angelica gigas, A. sinensis, and A. acutiloba have been considered as Angelicae radix in Korean, Chinese, and Japanese Pharmacopoeia, respectively. Since the origins of Angelicae radix differ from country to country, there is a need to develop an efficient analytical method to identify the origin of the Angelica root. In order to obtain chemical fingerprints, three different Angelicae Radices were analyzed by direct analysis in real time mass spectrometry (DART-MS). Significantly different DART-MS spectra were observed from three different species of Angelicae Radix. Strong peaks of decursin or decusinol angelate, and its dimer were exclusively found from A. gigas. Ligustilide and linoleic acid were detected as the major component from A. acutiloba. The strongest ligustilide peak was observed from A. sinensis. DART-MS fingerprinting is a promising method for the rapid identification and/or quality control of Angelicae Radix.

Quantification and Graphical Method for DNA Fingerprinting (유전자검사자료의 통계분석을 위한 수량화 및 그래프 방법)

  • 박미라
    • The Korean Journal of Applied Statistics
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    • v.15 no.1
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    • pp.85-105
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    • 2002
  • To explore the relationships among frequencies for sets of alleles, within or between loci, is one of the first analyses in population genetic study. The general question is whether the frequency of a set of alleles is the same as the product of each of the separate allele frequencies. For two alleles of a single locus, Hardy-Weinberg equilibrium is tested and for an allele from each of two loci, linkage disequilibrium is tested. However, it is more useful if we can quantify and graphically represent this information. In this study, we suggest graphical methods to find associations between alleles. We also analyze the STR data of Korean population as an illustration.

Proteomic Comparison of Gibberella moniliformis in Limited-Nitrogen (Fumonisin-Inducing) and Excess-Nitrogen (Fumonisin-Repressing) Conditions

  • Choi, Yoon-E;Butchko, Robert A.E.;Shim, Won-Bo
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.780-787
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    • 2012
  • The maize pathogen Gibberella moniliformis produces fumonisins, a group of mycotoxins associated with several disorders in animals and humans, including cancer. The current focus of our research is to understand the regulatory mechanisms involved in fumonisin biosynthesis. In this study, we employed a proteomics approach to identify novel genes involved in the fumonisin biosynthesis under nitrogen stress. The combination of genome sequence, mutant strains, EST database, microarrays, and proteomics offers an opportunity to advance our understanding of this process. We investigated the response of the G. moniliformis proteome in limited nitrogen (N0, fumonisin-inducing) and excess nitrogen (N+, fumonisin-repressing) conditions by one- and two-dimensional electrophoresis. We selected 11 differentially expressed proteins, six from limited nitrogen conditions and five from excess nitrogen conditions, and determined the sequences by peptide mass fingerprinting and MS/MS spectrophotometry. Subsequently, we identified the EST sequences corresponding to the proteins and studied their expression profiles in different culture conditions. Through the comparative analysis of gene and protein expression data, we identified three candidate genes for functional analysis and our results provided valuable clues regarding the regulatory mechanisms of fumonisin biosynthesis.