• Title/Summary/Keyword: Marker Selection

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Improvement of the Discrimination Capacity through the Expansion of Y Chromosomal STR Markers

  • Dong Gyu Lee;So Eun Lee;Ji Hwan Park;Si-Keun Lim;Ju Yeon Jung
    • Biomedical Science Letters
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    • v.29 no.4
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    • pp.302-313
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    • 2023
  • Y chromosomal short tandem repeat (Y-STR) markers have been developed continuously to complement forensic DNA analyses and population genetic studies. Initially, we collected data from previously reported Korean population Y-STR haplotype studies on 1133 individuals. We then conducted a marker expansion analysis using a dataset from the Y-STR Haplotype Reference Database (YHRD), covering up to 29 Y-STRs, referred to as Ymax. Additionally, we examined the impact of rapidly mutating (RM) Y-STRs included in this expanded marker set on the discrimination capacity. We observed that marker expansions both with (0.9896), and without (0.9510), RM Y-STR improved the discrimination capacity. Subsequently, we focused on 16 individuals belonging to seven distinct groups sharing identical haplotypes. These particular haplotypes had been previously identified among 476 unrelated males using 23 Y-STR markers from the PowerPlex® Y23 System. We expanded the marker panel up to Ymax to explore how discrimination improved with an expansion of Y-STR markers for these 16 individuals. Among the expanded markers, DYS627, which had high discriminatory power, had a high mutation rate (1.10 × 10-2) and high gene diversity (0.83). In contrast, DYF387S1 displayed high gene diversity (0.95) but a relatively low mutation rate (2.80 × 10-3). We propose that these findings will be valuable in the selection of suitable Y-STR markers, depending on the objectives of forensic analyses. Additionally, the presence of frequently observed Y-haplotypes in Korean population will facilitate statistical interpretation in Y-STR DNA profiling.

Assessment of Risks and Benefits of Using Antibiotics Resistance Genes in Mesenchymal Stem Cell-Based Ex-Vivo Therapy

  • Narayan Bashyal;Young Jun Lee;Jin-Hwa Jung;Min Gyeong Kim;Kwang-Wook Lee;Woo Sup Hwang;Sung-Soo Kim;Da-Young Chang;Haeyoung, Suh-Kim
    • International Journal of Stem Cells
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    • v.16 no.4
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    • pp.438-447
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    • 2023
  • Recently, ex-vivo gene therapy has emerged as a promising approach to enhance the therapeutic potential of mesenchymal stem cells (MSCs) by introducing functional genes in vitro. Here, we explored the need of using selection markers to increase the gene delivery efficiency and evaluated the potential risks associated with their use in the manufacturing process. We used MSCs/CD that carry the cytosine deaminase gene (CD) as a therapeutic gene and a puromycin resistance gene (PuroR) as a selection marker. We evaluated the correlation between the therapeutic efficacy and the purity of therapeutic MSCs/CD by examining their anti-cancer effect on co-cultured U87/GFP cells. To simulate in vivo horizontal transfer of the PuroR gene in vivo, we generated a puromycin-resistant E. coli (E. coli/PuroR) by introducing the PuroR gene and assessed its responsiveness to various antibiotics. We found that the anti-cancer effect of MSCs/CD was directly proportional to their purity, suggesting the crucial role of the PuroR gene in eliminating impure unmodified MSCs and enhancing the purity of MSCs/CD during the manufacturing process. Additionally, we found that clinically available antibiotics were effective in inhibiting the growth of hypothetical microorganism, E. coli/PuroR. In summary, our study highlights the potential benefits of using the PuroR gene as a selection marker to enhance the purity and efficacy of therapeutic cells in MSC-based gene therapy. Furthermore, our study suggests that the potential risk of horizontal transfer of antibiotics resistance genes in vivo can be effectively managed by clinically available antibiotics.

Identification of DNA Markers Related to Resistance to Herbicide Containing Mesotrione in Tongil Type Rice (통일형 벼에서 메소트리온계 제초제 저항성 연관 DNA marker 탐색)

  • Lee, Ji-Yoon;Cho, Jun-Hyeon;Lee, Jong-Hee;Cho, Su-Min;Kwon, Young-Ho;Park, Dong-Soo;Song, You-Chun;Ko, Jong-Min
    • Korean Journal of Breeding Science
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    • v.50 no.4
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    • pp.387-395
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    • 2018
  • This study was conducted to identify DNA markers related to resistance to herbicide containing mesotrione in Tongil type rice. Two Tongil type elite lines; Milyang154 and Suweon382, showed resistance to mesotrione, whereas the others were susceptible at 20 days after mesotrione application, and severe growth inhibition was observed in the remaining 13 lines. As a result of analysis of mesotrione resistance using 190 $F_2$ populations derived from a cross of Hanareum2 (susceptible) and Milyang154 (resistant), the mesotrione resistance locus was shown to be a single dominant gene with a 3:1 segregation ratio ($X^2=1.19$, P=0.31). To identify a DNA marker closely linked to the mesotrione resistance gene, bulked segregant analysis (BSA) was adopted. The DNA marker RM3501 was identified on chromosome 2 with a recombinant value of 0.53 to the mesotrione resistance gene. Mst1(t) was located between SSR (simple sequence repeat) markers RM3501 and RM324 with a physical map distance of 10.2 Mb-11.4 Mb on chromosome 2. The band pattern of agarose gel electrophoresis of the SSR marker RM3501 showed the same segregation pattern with respect to mesotrione treatment in 20 Tongil type varieties and a $BC_2F_2$ segregation population derived from a cross between Unkwang (resistant) and Hanareum2 (susceptible). Thus, the RM3501 DNA marker could be used in breeding programs for Marker Assisted Selection in mesotrione resistant rice breeding.

Asymmetry of Cotransduction Frequency Occured When the Transposon Is Used as a Marker (전위인자가 표식인자로 사용될때 발생하는 동시형질도입의 비대칭성)

  • 김창훈;박찬규
    • Korean Journal of Microbiology
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    • v.30 no.2
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    • pp.83-87
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    • 1992
  • When the transposon is used as a selectable marker. the cotransduction frequency depends on the selection order of the markers. In this study. we adopted a mathematical approach to explain this phenomenon. At first, the formation of transducing particles were considered in five different configurations. Then the probability functions indicating the possibilities for a marker to be fixed were mathematically formulated on the basis of probability density concept. After actual values and useful assumptions were integrated into the Formula. resulting frequencies from theoretical calculations were compared with actual data. Such analysis let us conclude that the asymmetry in the frequency arose from the lack of homology required for homologous recombination due to the transposon insertion and from the suppression of recombination around the region where the transposon is inserted.

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An Intelligent System of Marker Gene Selection for Classification of Cancers using Microarray Data (마이크로어레이 데이터를 이용한 암 분류 표지 유전자 선별 시스템)

  • Park, Su-Young;Jung, Chai-Yeoung
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.14 no.10
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    • pp.2365-2370
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    • 2010
  • The method of cancer classification based on microarray could contribute to being accurate cancer classification by finding differently expressing gene pattern statistically according to a cancer type. Therefore, the process to select a closely related informative gene with a particular cancer classification to classify cancer using present microarray technology with effect is essential. In this paper, the system can detect marker genes to likely express the most differentially explaining the effects of cancer using ovarian cancer microarray data. And it compare and analyze a performance of classification of the proposed system with it of established microarray system using multi-perceptron neural network layer. Microarray data set including marker gene that are selected using ANOVA method represent the highest classification accuracy of 98.61%, which show that it improve classification performance than established microarray system.

Improved LiDAR-Camera Calibration Using Marker Detection Based on 3D Plane Extraction

  • Yoo, Joong-Sun;Kim, Do-Hyeong;Kim, Gon-Woo
    • Journal of Electrical Engineering and Technology
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    • v.13 no.6
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    • pp.2530-2544
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    • 2018
  • In this paper, we propose an enhanced LiDAR-camera calibration method that extracts the marker plane from 3D point cloud information. In previous work, we estimated the straight line of each board to obtain the vertex. However, the errors in the point information in relation to the z axis were not considered. These errors are caused by the effects of user selection on the board border. Because of the nature of LiDAR, the point information is separated in the horizontal direction, causing the approximated model of the straight line to be erroneous. In the proposed work, we obtain each vertex by estimating a rectangle from a plane rather than obtaining a point from each straight line in order to obtain a vertex more precisely than the previous study. The advantage of using planes is that it is easier to select the area, and the most point information on the board is available. We demonstrated through experiments that the proposed method could be used to obtain more accurate results compared to the performance of the previous method.

Mapping of grain alkali digestion trait using a Cheongcheong/Nagdong doubled haploid population in rice

  • Kim, Hak Yoon;Kim, Kyung-Min
    • Journal of Plant Biotechnology
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    • v.43 no.1
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    • pp.76-81
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    • 2016
  • We performed a molecular marker-based analysis of quantitative trait loci for traits that determine the quality of appearance of grains using 120 doubled haploid lines developed by anther culture from the F1 cross between 'Cheongcheong' (Oryza sativa L. ssp. Indica) and 'Nagdong' (Oryza sativa L. ssp. Japonica). We therefore calculated the alkali digestion value (ADV), used to indirectly measure gelatinization temperature, to evaluate the quality of cooked rice in 2013 and 2014. The ADV score of frequency distribution was higher milled rice than brown rice. In total, nine different quantitative trait loci (QTLs) were found on 5 chromosomes in 2013 and 2014. Also, chromosome 5, 8 were detected over two years. We conclude that selected molecular markers from this QTL analysis could be exploited in future rice quality. In conclusion, we investigated ADV of brown and milled rice in CNDH population. This study found nine QTLs related to the ADV of brown and milled rice. The detected one marker can be used to select lines with desirable eating-quality traits because ADV is closely associated with the eating quality of cooked rice. Therefore, it will be useful to collect resources and distinguishable in many varieties for rice breeding program.

ISSR marker-assisted selection of male and female plants in a promising dioecious crop: jojoba (Simmondsia chinensis)

  • Sharma, Kuldeep;Agrawal, Veena;Gupta, Sarika;Kumar, Ravindra;Prasad, Manoj
    • Plant Biotechnology Reports
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    • v.2 no.4
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    • pp.239-243
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    • 2008
  • Simmondsia chinensis (Link) Schneider, a multipurpose and monogeneric dioecious shrub from arid zones, has emerged as a cash crop all over the globe. Its seed propagation poses severe problems due to its male-biased population: the male:female ratio is 5:1. Investigations have been carried out to generate a sex-specific Inter-simple sequence repeat (ISSR) marker for the early detection of male and female plants. Of the 42 primers analysed with a bulk sample of pooled male DNA and a bulk sample of pooled female DNA, only one primer, UBC-807, produced a unique ~1,200 base-pair fragment in the male DNA. To validate this observation, this primer was re-tested with individual male and female samples from eight cultivars. A similar unique ~1,200 bp fragment was present in the male individuals of all eight cultivars and completely absent in the female individuals tested. This is the first report of the use of ISSR markers to ascertain sex in physiologically mature S. chinensis plants.

An AFLP-based Linkage Map of Japanese Red Pine (Pinus densiflora) Using Haploid DNA Samples of Megagametophytes from a Single Maternal Tree

  • Kim, Yong-Yul;Choi, Hyung-Soon;Kang, Bum-Yong
    • Molecules and Cells
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    • v.20 no.2
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    • pp.201-209
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    • 2005
  • We have constructed an AFLP-based linkage map of Japanese red pine (Pinus densiflora Siebold et Zucc.) using haploid DNA samples of 96 megagametophytes from a single maternal tree, selection clone Kyungbuk 4. Twenty-eight primer pairs generated a total of 5,780 AFLP fragments. Five hundreds and thirteen fragments were verified as genetic markers with two alleles by their Mendelian segregation. At the linkage criteria LOD 4.0 and maximum recombination fraction 0.25(${\theta}$), a total of 152 markers constituted 25 framework maps for 19 major linkage groups. The maps spanned a total length of 2,341 cM with an average framework marker spacing of 18.4 cM. The estimated genome size was 2,662 cM. With an assumption of equal marker density, 82.2% of the estimated genome would be within 10 cM of one of the 230 linked markers, and 68.1% would be within 10 cM of one of the 152 framework markers. We evaluated map completeness in terms of LOD value, marker density, genome length, and map coverage. The resulting map will provide crucial information for future genomic studies of the Japanese red pine, in particular for QTL mapping of economically important breeding target traits.

Transformation of a Filamentous Fungus Cryphonectria parasitica Using Agrobacterium tumefaciens

  • Park, Seung-Moon;Kim, Dae-Hyuk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.3
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    • pp.217-222
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    • 2004
  • As Agrobacterium tumefaciens, which has long been used to transform plants, is known to transfer T-DNA to budding yeast, Saccharomyces cerevisiae, a variety of fungi were subjected to the A. tumefaciens-mediated transformation to improve their transformation frequency and feasibility. The A. tumefaciens-mediated transformation of chestnut blight fungus, Cryphonectria parasitica, is performed in this study as the first example of transformation of a hardwood fungal pathogen. The transfer of the binary vector pBIN9-Hg, containing the bacterial hygromycin B phosphotransferase gene under the control of the Aspergillus nidulans trpC promoter and terminator, as a selectable marker, led to the selection of more than 1,000 stable, hygromycin B-resistant transformants per 1${\times}$10$\^$6/ conidia of C. parasitica. The putative transformants appeared to be mitotically stable. The transformation efficiency appears to depend on the bacterial strain, age of the bacteria cell culture and ratio of fungal spores to bacterial cells. PCR and Southern blot analysis indicated that the marker gene was inserted at different chromosomal sites. Moreover, three transformants out of ten showed more than two hybridizing bands, suggesting more than two copies of the inserted marker gene are not uncommon.