• Title/Summary/Keyword: MS markers

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Comparison of Plasma Proteome Expression between the Young and Mature Adult Pigs

  • Jeong, Jin Young;Nam, Jin Sun;Kim, Jang Mi;Jeong, Hak Jae;Kim, Kyung Woon;Lee, Hyun-Jeong
    • Reproductive and Developmental Biology
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    • v.37 no.4
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    • pp.247-253
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    • 2013
  • Here, we present an approach of blood plasma proteome profiling and their comparisons between the young and the adult pigs as prerequisite for the identification of bio-markers related to the health conditions, growth performance and meat quality. To profile the proteome in porcine plasma, blood samples were collected from 19 young piglets and 20 adult male barrows and the plasma was retrieved. Then, protein profiling was initiated using one and two-dimensional electrophoresis. Proteins were spotted and then identified by MALDI-TOF-TOF and LC-MS-MS. In the results, more than thirty-six and twenty eight protein spots were selected in young piglets and adult pigs, respectively and twenty three proteins were identified. The proteome profile images were compared between those ones using Image Master Version 7.0. The image of expressed proteome showed that most of proteins from plasma of young piglet separated clearly and concentrated in 2DE display compared to ones from adult. Image analysis in detail was carried out to look for the specific proteins related to age progression. It demonstrated that the characteristics of proteome expression could be distinct to their age stages. Further investigations needed to proceed to understand the age dependent change of protein conformation and biological meaning of those differences in proteome expression between young and mature adult pigs.

Discrimination of biological and artificial nicotine in e-liquid

  • Hyoung-Joon Park;Heesung Moon;Min Kyoung Lee;Min Soo Kim;Seok Heo;Chang-Yong Yoon;Sunyoung Baek
    • Analytical Science and Technology
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    • v.36 no.1
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    • pp.22-31
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    • 2023
  • As the use of e-liquid cigarettes is rapidly increasing worldwide, it multiplies the potential risk undisclosed to the health of non- and smokers. To reduce the hazard, each country has its own set of regulations for controlling e-liquids. In Korea, the narrow definition of tobacco makes it difficult and have been steadily occurring tax evasion exploiting the difference in natural and artificial nicotine. Therefore, it is very important to distinguish source of nicotine for their regulation. To find biochemical discriminant markers, this study established analysis methods based on high-performance liquid chromatography coupled with diode array detector (HPLC-DAD) and high-performance liquid chromatography coupled with triple Quadrupole mass spectrometry (HPLC-MS/MS) for nicotine enantiomers and tobacco alkaloids targeted using the difference in pathways of nicotine biosynthesis and chemical synthesis. The method was validated by experimenting linearity (R2 > 0.999), recovery (80.99-108.41 %), accuracy (94.11-109.73 %) and precision (0.04-8.27 %). Then, the results for discrimination of the nicotine obtained from analysis of 65 commercial e-liquid products available in Korean market was evaluated. The method successfully applied to the e-liquids and one sample labelled 'synthetic nicotine' for tax exemption was found to contain a natural nicotine product. This method can be used to determine whether an e-liquid product uses natural or artificial nicotine and monitor non-taxable e-liquid products. The method is more scientific than the existing one, which relies only on field evidence.

Detection of Radiation-Induced Hydrocarbons in Green, Black and Oolong Teas

  • Kausar, Tusneen;Lee, Jung-Eun;Noh, Jung-Eun;Kwon, Young-Ju;Kwon, Joong-Ho
    • Preventive Nutrition and Food Science
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    • v.9 no.3
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    • pp.222-226
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    • 2004
  • Hydrocarbons induced by gamma-irradiation of green, black, and oolong teas were analyzed to determine whether the hydrocarbons can be used as markers for detecting post-irradiation of these teas. The samples were irradiated at 0, 2.5, 5, 7.5, and 10 kGy. Detection was attempted by extracting fat from the teas, separation of hydrocarbons with florisil column chromatography, and identification of hydrocarbons by gas chromatography-mass spectroscopy (GC-MS). Concentration of hydrocarbons increased with the irradiation dose. The major hydrocarbons in irradiated green, black, and oolong teas were 1-tetradecence (14:1), pentadecane (15:0), 1,7-hexadecadiene (16:2), 1-hexadecene (16:1), 8-heptadecene (17:1), and heptadecane (17:0). Radiation-induced hydrocarbons in teas were 1,7-hexadecadiene and 8-heptadecene. These compounds were not detected in non-irradiated samples, so the hydrocarbons (16:2, 17:1) can be used as markers for detecting post-irradiation of the teas. Furthermore, detection of hydrocarbons after 12 months storage at room temperature remains a suitable method for identifying irradiated teas.

Verification of ET and AI Derived Offspring Using on the Genetic Polymorphisms of Microsatellite and Coat Color Related Genes in Jeju Black Cattle (제주흑우 집단에서 모색 관련 유전자와 microsatellite marker의 다형현상을 이용한 수정란이식 및 인공수정 유래 후대우 검증)

  • Han, Sang-Hyun;Ko, Jin-Cheul;Kim, Young-Hoon;Kim, Nam-Young;Kim, Jae-Hwan;Ko, Moon-Suck;Jeong, Ha-Yeon;Cho, In-Cheol;Yang, Young-Hoon;Lee, Sung-Soo
    • Journal of Life Science
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    • v.20 no.3
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    • pp.381-387
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    • 2010
  • To find offspring of Jeju Black cattle (JBC) produced by embryo transfer (ET) and artificial insemination (AI), a molecular genetic study was carried out in candidate cattle populations collected from cattle farms in Jeju Island, Korea. The genetic marker set was composed of 11 ISAG microsatellite (MS) markers, 11 SAES MS markers selected by our preliminary analysis for population diversity of JBC and two major coat color related genes: MC1R and ASIP. The results showed a combined non-exclusion probability for first parent (NE-P1) that was higher than that recommended by ISAG (above 0.9995), and a combined non-exclusion probability for sib identity of $5.3{\times}10^{-10}$. Parentage analysis showed that the cases identified the candidate's father only (77.0%), mother only (54.0%), and both parents (40.5%) in the candidate offspring population. The ET and AI calves were identified as 14.7% in the in vitro fertilized eggs provided and 32.4% in total population, respectively. However, the result from ISAG marker analysis showed 3 identical allele-combinations in 7 calves, and that from ISAG/SAES MS marker combination also showed 1 identical allele-combination in 2 calves. Data from MS and coat-color gene analyses provided information for complete identification of all animals tested. Because the present JBC population was mostly bred using small nuclear founders through bioengineering techniques such as AI and ET, the genetic diversity levels obtained from MS analysis in the JBC population were relatively lower than those of other cattle populations, including Hanwoo. The results suggested that the more efficient marker combinations, including coat color related genotypes, should be studied and used for constructing a system for identification and molecular breeding of JBC as well.

In vitro micropropagation of M.26 (Malus pumila Mill) apple rootstock and assessment of the genetic diversity of proliferated plantlets using simple sequence repeat markers (사과 대목 M.26 (Malus pumila Mill)의 기내 대량번식 및 simple sequence repeat 마커를 이용한 증식된 식물체의 유전적 다양성 평가)

  • Cho, Kang Hee;Han, Bong Hee;Han, Jeom Hwa;Park, Seo Jun;Kim, Se Hee;Lee, Han Chan;Kim, Mi Young;Kim, Myung-Su
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.382-391
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    • 2018
  • The objective of this study was to determine the most effective medium condition of shoot proliferation and root formation for the efficient in vitro micropropagation of M.26 (Malus pumila Mill). Simple sequence repeat (SSR) markers were used to analyze the genetic diversity of micro-propagated and greenhouse grown M.26. Shoot proliferation was carried out in MS (Murashige and Skoog) containing benzyladenin (BA, $0.5{\sim}5.0mg{\cdot}L^{-1}$) and thidiazuron (TDZ, $0.01{\sim}0.1mg{\cdot}L^{-1}$). The highest number of shoots (10.67 shoots per explant) was induced by adding BA at a concentration $1.0mg{\cdot}L^{-1}$. TDZ treatments caused higher hyperhydricity rate in cultured explants than in BA treatments. There was no significant effect of both BA and auxin on shoot proliferation, and the optimum proliferation medium for M.26 was MS medium containing $1.0mg{\cdot}L^{-1}$ BA. To find a suitable medium composition for shoot rooting, we tested different concentrations indole-3-butyric acid (IBA) and ${\alpha}$-naphthaleneacetic acid ($0.5{\sim}5.0mg{\cdot}L^{-1}$), MS medium (1/4-1), sucrose ($0{\sim}30g{\cdot}L^{-1}$). The shoots showed good rooting on half-strength MS medium containing $1.0mg{\cdot}L^{-1}$ IBA and $15-20g{\cdot}L^{-1}$ sucrose. The rooting rate (100%), number of roots (10.45 ~ 13.60 roots per explant), root length (7.41 ~ 8.33 cm), and shoot length (4.93 ~ 5.38 cm) were good on this medium. Fifteen SSR primers were detected in a total of 30 alleles in 20 micro-propagated plantlets, all SSR profiles from micro-propagated plantlets were monomorphic and similar to greenhouse grown control plantlet M.26 plant. The results indicated that M.26 micro-propagated plantlets were genetically stable.

Comparison for Genetic Diversity between Korean Native Commercial Chicken Brand Groups using Microsatellite Markers (Microsatellite Marker를 활용한 토종닭 브랜드 집단 간의 유전적 다양성 분석)

  • Lee, Hak-Kyo;Oh, Jae-Don;Park, Chan-Ho;Lee, Kun-Woo;Lee, Jun-Heon;Jeon, Gwang-Joo;Kong, Hong-Sik
    • Korean Journal of Poultry Science
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    • v.37 no.4
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    • pp.355-360
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    • 2010
  • To estimate the genetic characteristics within two brands of Korean native commercial chicken, we used a total of 302 genomic DNAs from two groups (Woorichicken: 152, Hanhyup3chicken: 150). Sizes of 10 microsatellite markers were decided using GeneMapper Software (v.4.0) after analyzing ABI 3130. Genetic diversity indices including expected heterozygosity (Ex H), observed heterozygosity (Ob H) and polymorphism information content (PIC). Frequencies of microsatellites markers were used to estimate heterozygosities and genetic distances. LEI0073 showed the highest value in all genetic diversity (Ex H, Ob H and PIC). On the other hand, MCW322 showed the lowest value in all genetic diversity. The calculated genetic distance of the two brand groups is 0.199 (standard genetic distance) and 0.132 (DA distance). Genetic distances of the two groups were relatively close to each other. Each individual is ramified to two brand groups in phylogenetic dendrogram.

Hybridity Verification of Progenies Obtained from Ovule Culture by Using RAPD Markers in Reciprocal Crosses of Alstroemeria (알스트로메리아 배주배양을 통하여 획득한 정역교배 자손의 혼종성 분석)

  • Lee, Ja-Hyun;Joung, Youn-Hwa;Han, Tae-Ho
    • FLOWER RESEARCH JOURNAL
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    • v.19 no.4
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    • pp.231-237
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    • 2011
  • In this study, we performed ovule culture after reciprocal crosses of two Alstroemeria accessions and investigated genetic contribution of parents by using RAPD markers. The best method was half-ovule culture on MS medium supplemented with $60g{\cdot}L^{-1}$ sucrose and $2.2g{\cdot}L^{-1}$ gelrite at 14 days after pollination. Embryos began to germinate after 6 weeks of culture. The complete plantlets were formed after 4 months of culture. In eight progenies and two parental cultivars, 59 polymorphic bands were obtained out of 89 total bands by RAPD analysis using 7 primers. Eight $F_1$ progenies from the crosses between two accessions using reciprocal crosses showed 1:1 contribution of maternal and paternal parents. It is confirmed that $F_1$ progenies were obtained from parental accessions by using RAPD markers. We conclude this cross combination showed pre-fertilization barriers with incompatibility between stigma or style, and pollen because progeny number was different in each cross combination. Thereby, it warrants overcoming pre-fertilization barrier together with post-fertilization barrier in order to broaden the heterozygosity within progeny populations in Alstroemeria breeding program.

Time-Dependent Hepatic Proteome Analysis in Lean and Diet-Induced Obese Mice

  • Oh, Tae-Seok;Kwon, Eun-Young;Choi, Jung-Won;Choi, Myung-Sook;Yun, Jong-Won
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1211-1227
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    • 2011
  • C57BL/6J mice have been widely used as a diet-induced obesity model because they trigger common features of the human metabolic syndrome. In the present study, C57BL/6J male mice were fed either a high-fat diet (HFD) or normal diet (ND) during a 24-week period, and then the age-dependent liver proteome of mice in two groups was analyzed using 2-DE combined with MALDI-TOF-MS. Among identified proteins, up-regulated proteins were subdivided to early (during the first 4 weeks) and late (20~24 weeks) markers that played a role in diet-induced obesity development. Important early markers included ketohexokinase and prohibitin, and late markers included the 75 kDa glucose-regulated protein, citrate synthase, and selenium-binding liver protein. Of these, the 75 kDa glucosere-gulated protein has already been linked to obesity; however, prohibitin protein involved in obesity was identified for the first time in this study. In order to validate the proteomic results and gain insight into metabolic changes between the two groups, we further confirmed the expression pattern of some proteins of interest by Western blot analysis. Combined results of proteomic analysis with Western blot analysis revealed that antioxidant enzymes were progressively decreased, whereas cytoskeletal proteins were time-dependently increased in HFD mice.

Evaluation of BTA1 and BTA5 QTL Regions for Growth and Carcass Traits in American and Korean Cattle

  • Kim, K.S.;Kim, S.W.;Raney, N.E.;Ernst, C.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.11
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    • pp.1521-1528
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    • 2012
  • Previously identified QTL regions on BTA1 and BTA5 were investigated to validate the QTL regions and to identify candidate genes for growth and carcass traits in commercial cattle populations from the USA and Korea. Initially, a total of 8 polymorphic microsatellite (MS) markers in the BTA1 and 5 QTL regions were used for Chi-square tests to compare the frequencies of individual alleles between high and low phenotypic groups for the US (Michigan Cattleman's Association/Michigan State University; MCA/MSU) cattle. For a subsequent study, 24 candidate genes containing missense mutations and located within the QTL regions based on bovine genome sequence data were analyzed for genotyping in the two commercial cattle populations. Re-sequencing analyses confirmed 18 public missense SNPs and identified 9 new SNPs. Seventeen of these SNPs were used for genotyping of the MCA/MSU cattle (n = 98) and Korean native cattle (n = 323). On BTA1, UPK1B, HRG, and MAGEF1 polymorphisms residing between BM1312 and BMS4048 were significantly associated with growth and carcass traits in one or both of the MCA/MSU and Korean populations. On BTA5, ABCD2, IL22 and SNRPF polymorphisms residing between BL4 and BR2936 were associated with marbling and backfat traits in one or both of the MCA/MSU and Korean cattle populations. These results suggested that BTA 1 and 5 QTL regions may be segregating in both Korean Hanwoo and USA commercial cattle populations and DNA markers tested in this study may contribute to the identification of positional candidate genes for marker-assisted selection programs.

Genotype Effect on Somatic Embryogenesis and Plant Regeneration of 15 Aralia elata (두릅나무 15개체의 체세포배 유도 및 식물체 재분화에 미치는 유전자형의 효과)

  • 문흥규;홍용표;김용욱;이재순
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.3
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    • pp.129-134
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    • 2001
  • Winter bud explants from 15 individual angelica tree (Aralia elata) were cultured in vitro to find out optimal conditions for somatic embryo induction as well as plant regeneration. Calli are induced and grown on MS medium supplemented with 1.0 mg/L 2,4-D for 4 weeks and subcultured on a half-strength MS medium without phytohormones to induce somatic embryos. Inter-simple sequence repeat (I-SSR) markers were analyzed with total DNAs extracted from the trees. Genotype effects on somatic embryo induction were examined by cluster analysis. Callus induction rate varied from 58.5 to 100% among the genotypes. Somatic embryo induction rate also greatly varied from 0 to 100% among the genotypes. There was a significant difference in somatic embryo induction rate even among the individual trees that showed close genetic relationships each other. This suggested that somatic embryo induction rate in Aralia elata be influenced by a few major specific genes rather than whole genomic similarity among individual trees. Four individuals of Ulneong-7, Cheju-1, Shingu and China, which are recalcitrant to somatic embryo induction, turned out to have a close genetic relationship, suggesting that both physiological and genetic factors affect somatic embryo induction. The results suggest that genotype selection be the most important factor to achieve an efficient propagation, although cultural optimization through medium and explant manipulation may also play crucial roles in somatic embryogensis as well as plant regeneration of these species.

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