• Title/Summary/Keyword: Data Amplification

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Application of Disease Resistance Markers for Developing Elite Tomato Varieties and Lines

  • Kim, Hyoun-Joung;Lee, Heung-Ryul;Hyun, Ji-Young;Won, Dong-Chan;Hong, Dong-Oh;Cho, Hwa-Jin;Lee, Kyung-Ah;Her, Nam-Han;Lee, Jang-Ha;Harn, Chee-Hark
    • Horticultural Science & Technology
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    • v.29 no.4
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    • pp.336-344
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    • 2011
  • Using the abundant available information about the tomato genome, we developed DNA markers that are linked to disease resistant loci and performed marker-assisted selection (MAS) to construct multi-disease resistant lines and varieties. Resistance markers of Ty-1, T2, and I2, which are linked to disease resistance to Tomato yellow leaf curl virus (TYLCV), Tomato mosaic virus (ToMV), and Fusarium wilt, respectively, were developed in a co-dominant fashion. DNA sequences near the resistance loci of TYLCV, ToMV, and Fusarium wilt were used for primer design. Reported candidate markers for powdery mildew-resistance were screened and the 32.5Cla marker was selected. All four markers (Ty-1, T2, I2, and 32.5Cla) were converted to cleavage amplification polymorphisms (CAPS) markers. Then, the CAPS markers were applied to 96 tomato lines to determine the phenetic relationships among the lines. This information yielded clusters of breeding lines illustrating the distribution of resistant and susceptible characters among lines. These data were utilized further in a MAS program for several generations, and a total of ten varieties and ten inbred lines were constructed. Among four traits, three were introduced to develop varieties and breeding lines through the MAS program; several cultivars possessed up to seven disease resistant traits. These resistant trait-related markers that were developed for the tomato MAS program could be used to select early stage seedlings, saving time and cost, and to construct multi-disease resistant lines and varieties.

Relationship between Cell Size and Specific Thrombopoietin Productivity in Chinese Hamster Ovary Cells during Dihydrofolate Reductase-mediated Gene Amplification

  • Kim, Tae-Kyung;Chung, Joo-Young;Sung, Yun-Hee;Lee, Gyun-Min
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.5
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    • pp.332-336
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    • 2001
  • When parental Chinese hamster ovary (CHO) cell clones that are capable of producing thrombopoietin (TPO) were subjected to high methotrexate (MTX) concentrations, clonal variations in cell growth were apparent. In the clones that had no significant enhancement in specific TPO productivity (q$\_$Tpo/)when a higher level of MTX was administered, their growth was not depressed significantly nor their cell size changed significantly. On the other hand, those clones that showed a significant-enhancement in q$\_$Tpo/ at higher a MTX dosage, cell growth was depressed initially but recovered during successive sub-cultures. Furthermore, their cell size increased, which suggested that changes in cell size may be indicative of an enhanced q$\_$Tpo/. When the enhancement of the q$\_$Tpo/ of 9 clones after a high MTX dosage was plotted against the extent of the increase of their size, there was a linear correlation (γ$^2$=0.80, p<0.001, ANOVA), which suggested that an enhancement of q$\_$Tpo/ after high MTX administration can be measured by the increase in their cell size. Taken together, our data demonstrate that the selection of amplified CHO cell clones with enhanced q$\_$Tpo/ can be done upon their increased size and growth pattern. This facilitates the development of highly productive recombinant CHO cell lines.

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Association between Motilin Receptor Gene Haplotypes and Growth Traits in Japanese Hinai-dori Crossbred Chickens

  • Takahashi, Hideaki;Rikimaru, Kazuhiro;Komatsu, Megumi;Uemoto, Yoshinobu;Suzuki, Keiichi
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.3
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    • pp.316-323
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    • 2014
  • We previously identified quantitative trait loci (QTL) for body weight and average daily gain in a common region between ADL0198 (chr 1: 171.7 Mb) and ABR0287 (chr 1: 173.4 Mb) on chicken chromosome 1 in an $F_2$ resource population produced by crossing low- and high-growth lines of the Hinai-dori breed. Motilin receptor (MLNR) is a candidate gene affecting growth traits in the region. In this study, we genotyped polymorphisms of the MLNR gene and investigated its association with growth traits in a Hinai-dori $F_2$ intercross population. All the exons of the MLNR gene in the parental population were subjected to PCR amplification, nucleotide sequenced and haplotypes identified. To distinguish resultant diplotype individuals in the $F_2$ population, a mismatch amplification mutation assay was performed. Three haplotypes (Haplotypes 1-3) were accordingly identified. Six genotypes produced by the combination of three haplotypes (Haplotype 1, 2, and 3) were examined in order to identify associations between MLNR haplotypes and growth traits. The data showed that Haplotype 1 was superior to Haplotype 2 and 3 in body weight at 10 and 14 weeks of age, average daily gain between 4 and 10 weeks, 10 and 14 weeks, and 0 and 14 weeks of age in female in $F_2$ females. It was concluded that MLNR is a useful marker of growth traits and could be used to develop strategies for improving growth traits in the Hinai-dori breed.

Molecular Identification of Korean Mountain Ginseng Using an Amplification Refractory Mutation System (ARMS)

  • In, Jun-Gyo;Kim, Min-Kyeoung;Lee, Ok-Ran;Kim, Yu-Jin;Lee, Beom-Soo;Kim, Se-Young;Kwon, Woo-Seang;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.34 no.1
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    • pp.41-46
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    • 2010
  • Expensive herbs such as ginseng are always a possible target for fraudulent labeling. New mountain ginseng strains have occasionally been found deep within mountain areas and commercially traded at exorbitant prices. However, until now, no scientific basis has existed to distinguish such ginseng from commonly cultivated ginseng species other than by virtue of being found within deep mountain areas. Polymerase chain reaction (PCR) analysis of the internal transcribed spacer has been shown to be an appropriate method for the identification of the most popular species (Panax ginseng) in the Panax ginseng genus. A single nucleotide polymorphism (SNP) has been identified between three newly found mountain ginseng (KGD4, KGD5, and KW1) and already established Panax species. Specific PCR primers were designed from this SNP site within the sequence data and used to detect the mountain ginseng strains via multiplex PCR. The established multiplex-PCR method for the simultaneous detection of newly found mountain ginseng strains, Korean ginseng, and foreign ginseng in a single reaction was determined to be effective. This study is the first report of scientific discrimination of "mountain ginsengs" and describes an effective method of identification for fraud prevention and for uncovering the possible presence of other, cheaper ginseng species on the market.

Development of Smart Phone Application with User Fitting for Hearing Impaired in Various Environment (환경별 사용자 피팅이 가능한 스마트폰 보청기 어플리케이션 개발)

  • Lee, K.;Kang, S.-I.;Park, G.S.;Lee, S.M.
    • Journal of rehabilitation welfare engineering & assistive technology
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    • v.9 no.2
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    • pp.121-128
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    • 2015
  • In this paper, we introduce our smart phone application for hearing impaired people. Unlike previous hearing aids application, our application has both functions of self hearing tests and sound amplification. We provide user with two types of self hearing test, those are standard hearing test and detail hearing test. Each test has different advantages like short test time and accurate test result, however both tests provide reliable result. Our hearing test allows that user can measure their hearing loss level according to the frequency. Sound amplification function provide proper fitting formula by using NAL-NL1, and it guarantees high speech intelligibility. Our application can also memorize user's fitting data at specific test condition. Therefore those who using our hearing aids application can easily reset their hearing aids / earphone properties periodically. Our application is advanced at the aspect of self fitting function that no other previous application did. So we are expecting these comfortable processes about fitting lead the number of hearing aids user to be increased.

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Immunohistochemical Detection of N-myc Gene Product by Using Antiserum Against Synthetic Peptide (항-펩타이드 항체를 이용한 암유전자 N-myc 산물의 면역조직화학적 검출)

  • Lee, Hyun-Chul;Lee, Wan-Joo;Ahn, Tai-Hew
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.2
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    • pp.167-174
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    • 1987
  • N-myc, a DNA sequence related to the oncogene c-myc, was found to be amplified in untreated primary neuroblastomas and the amplification appeared to be associated with advanced disease at diagnosis and rapid tumor progression. Synthetic peptides have been useful immunogens for generating antisera and monoclonal antibodies to a number of native proteins. In order to identify myc-related protein in the tumor cells, an antiserum against a synthetic hexapeptide (-Glu-Asp-Ile-Trp-Lys-Lys-), whose sequence corresponds to a part of the exon 2 of oncogene N-myc, was prepared by immunizing a rabbit with BSA-conjugated peptide. After ammonium sulfate precipitation and affinity column chromatography, it appeared to be specific to the peptide. Strong nuclear staining in immunoperoxidase method using this serum was observed in both human promyeloid leukemic cell line, HL-60(containing high c-myc copy number), and human neuroblastoma cell line, LA-N-5 (containing high N-myc copy number), whereas LA351 (human lymphoid cell line) cells did not react with the serum. This reaction was completely abrogated by incubating the antiserum with soluble excess peptide. These data suggest that the protein encoded by N-myc could be localized in the nucleus as c-myc protein and this antiserum can be used to detect myc-related tumor cells in clinical samples and to determine if the N-myc expression correlates with genomic amplification in cell lines, untreated primary tumors, and untreated metastases.

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Molecular identification of Allium ochotense and Allium microdictyon using multiplex-PCR based on single nucleotide polymorphisms

  • Kim, Yong-Bog;Ramekar, Rahul Vasudeo;Choi, Seong-Jin;Choi, Byoung-Gon;Kim, Se-Won;Moon, Youn-Ki;Noh, Hee-Sun;Lee, Ju-Kyong;Hong, Jin-Sung;Park, Nam-Il;Choi, Ik-Young;Choi, Seon-Kang;Park, Kyong-Cheul
    • Horticulture, Environment, and Biotechnology : HEB
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    • v.59 no.6
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    • pp.865-873
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    • 2018
  • Allium ochotense and Allium microdictyon are commonly known as 'Mountain garlic' and are popular, economically important species in many countries such as Korea, China, and Mongolia. Their leaves are used as culinary side dishes and in traditional medicines. In Korea, these two species are at risk of extinction due to damage to their natural habitat and thus, conservation and breeding programs are needed. However, their identification relies mostly on morphological data, which is limited and until recently, led to classifying these two species under A. victorialis. In the present study, a simple and reliable method of molecular identification was developed to distinguish A. ochotense from A. microdictyon that targets four barcoding regions: the internal transcribed spacer (ITS), the maturase K gene (matK), the chloroplast psbA-trnH intergenic region, and the ribulose-bisphosphate carboxylase large subunit gene (rbcL). Single nucleotide polymorphisms (SNPs) were found in ITS and matK regions, and species-specific primers were designed based solely on the SNP at position 680 of the ITS region that could differentiate A. ochotense from A. microdictyon. Using these primers in amplification refractory mutation system (ARMS)-PCR, A. ochotense, and A. microdictyon could be simultaneously and efficiently distinguished. This study is the first to report a simple, rapid, and efficient method for discriminating A. ochotense and A. microdictyon, indicating the utility of species-specific markers in the development of conservation and breeding programs.

A comparative study of the prevalence of Helicobacter pylori in the oral biofilms of a group of dental and non-dental undergraduates from Sri Lanka

  • Mallikaarachchi, MADKS;Rajapakse, Sanath;Gunawardhana, KSND;Jayatilake, JAMS
    • International Journal of Oral Biology
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    • v.46 no.1
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    • pp.60-65
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    • 2021
  • Dental health care workers (DHCW) are at a risk of occupational exposure to Helicobacter pylori from the aerosolized oral biofilms and saliva of patients. We designed this study to investigate the prevalence of H. pylori in the oral biofilms of a group of dental and non-dental undergraduates from Sri Lanka. After obtaining informed consent, oral biofilms were collected from 38 dental undergraduates (19 males and 19 females) undergoing clinical training and 33 non-dental undergraduates (14 males and 19 females). The participants were in the age range of 22-27 years and had healthy periodontium. Total DNA from the oral biofilms were extracted, and H. pylori DNA was detected using polymerase chain reaction (PCR) amplification of 16S rRNA gene of H. pylori using JW22-JW23 primers, and the results were confirmed using PCR amplification of H. pylori-urease specific HPU1-HPU2 primers. Out of 71 participants, 11 (28.95%) dental and 3 (9.09%) non-dental undergraduates had H. pylori in their oral biofilms indicating an overall prevalence rate of 19.72% (14/71). Thus, the prevalence of H. pylori in oral biofilms was significantly higher in dental undergraduates than in non-dental undergraduates (p < 0.05). An odds ratio of 4.07 indicated that dental undergraduates were four times more likely to harbor H. pylori in their oral biofilms than non-dental undergraduates. Foregoing data support the fact that there may be greater occupational risk of exposure to H. pylori for dental undergraduates during clinical training than that for non-dental undergraduates, warranting meticulous infection control practices during clinical dentistry.

Combination of multiplex reverse transcription recombinase polymerase amplification assay and capillary electrophoresis provides high sensitive and high-throughput simultaneous detection of avian influenza virus subtypes

  • Tsai, Shou-Kuan;Chen, Chen-Chih;Lin, Han-Jia;Lin, Han-You;Chen, Ting-Tzu;Wang, Lih-Chiann
    • Journal of Veterinary Science
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    • v.21 no.2
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    • pp.24.1-24.11
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    • 2020
  • The pandemic of avian influenza viruses (AIVs) in Asia has caused enormous economic loss in poultry industry and human health threat, especially clade 2.3.4.4 H5 and H7 subtypes in recent years. The endemic chicken H6 virus in Taiwan has also brought about human and dog infections. Since wild waterfowls is the major AIV reservoir, it is important to monitor the diversified subtypes in wildfowl flocks in early stage to prevent viral reassortment and transmission. To develop a more efficient and sensitive approach is a key issue in epidemic control. In this study, we integrate multiplex reverse transcription recombinase polymerase amplification (RT-RPA) and capillary electrophoresis (CE) for high-throughput detection and differentiation of AIVs in wild waterfowls in Taiwan. Four viral genes were detected simultaneously, including nucleoprotein (NP) gene of all AIVs, hemagglutinin (HA) gene of clade 2.3.4.4 H5, H6 and H7 subtypes. The detection limit of the developed detection system could achieve as low as one copy number for each of the four viral gene targets. Sixty wild waterfowl field samples were tested and all of the four gene signals were unambiguously identified within 6 h, including the initial sample processing and the final CE data analysis. The results indicated that multiplex RT-RPA combined with CE was an excellent alternative for instant simultaneous AIV detection and subtype differentiation. The high efficiency and sensitivity of the proposed method could greatly assist in wild bird monitoring and epidemic control of poultry.

Frequency Analysis of Extreme High Water Level Considering Tide/Surge Characteristic Changes (조석/해일 환경병화를 감안한 고극조위 빈도분석)

  • Kang, Ju Whan;Moon, Seung Rok;Park, Seon Jung
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.26 no.1B
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    • pp.99-106
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    • 2006
  • Due to both global warming and constructions of seadike/seawalls, continuously or abruptly rising tendencies of extreme high water levels have been detected at Kunsan and Mokpo harbors. This paper deals firstly with the separation of each effect, namely global warming effect and construction effect, on increases of water level quantitatively by a linear regression method. And then, it can be explained why and how the extreme high water levels had been risen just after constructions at both harbors. A numerical simulation of $M_2$ tidal constituent at Mokpo coastal zone shows that the tidal amplification by constructions is mainly due to the extinguishment of TCE at Mokpogu. The tidal flat effect makes the amplification more deepen at spring tide or extreme high tide, which results in the increase of inundation risk at Mokpo harbor. A frequency analysis method is applied, which is shown to be effective at such a site of having non-homogeneous tidal data due to constructions as Mokpo harbor.