• Title/Summary/Keyword: Marker selection

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Development of a Molecular Selection Marker for Bacillus licheniformis K12 (Bacillus licheniformis K12 균주 분자 선발 마커 개발)

  • Young Jin Kim;Sam Woong Kim;Tae Wok Lee;Won-Jae Chi;Woo Young Bang;Ki Hwan Moon;Tae Wan Kim;Kyu Ho Bang;Sang Wan Gal
    • Journal of Life Science
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    • v.33 no.10
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    • pp.808-819
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    • 2023
  • This study was conducted to develop a selection marker for the identification of the Bacillus licheniformis K12 strain in microbial communities. The strain not only demonstrates good growth at moderate temperatures but also contains enzymes that catalyze the decomposition of various polymer materials, such as proteases, amylases, cellulases, lipases, and xylanases. To identify molecular markers appropriate for use in a microbial community, a search was conducted to identify variable gene regions that show considerable genetic mutations, such as recombinase, integration, and transposase sites, as well as phase-related genes. As a result, five areas were identified that have potential as selection markers. The candidate markers were two recombinase sites (BLK1 and BLK2), two integration sites (BLK3 and BLK4), and one phase-related site (BLK5). A PCR analysis performed with different Bacillus species (e.g., B. licheniformis, Bacillus velezensis, Bacillus subtilis, and Bacillus cereus) confirmed that PCR products appeared at specific locations in B. licheniformis: BLK1 in recombinase, BLK2 in recombinase family protein, and BLK3 and BLK4 as site-specific integrations. In addition, BLK1 and BLK3 were identified as good candidate markers via a PCR analysis performed on subspecies of standard B. licheniformis strains. Therefore, the findings suggest that BLK1 can be used as a selection marker for B. licheniformis species and subspecies in the microbiome.

Refinements for the amplification and sequencing of red algal DNA barcode and RedToL phylogenetic markers: a summary of current primers, profiles and strategies

  • Saunders, Gary W.;Moore, Tanya E.
    • ALGAE
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    • v.28 no.1
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    • pp.31-43
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    • 2013
  • This review provides a comprehensive summary of the PCR primers and profiles currently in use in our laboratory for red algal DNA barcoding and phylogenetic research. While work focuses on florideophyte taxa, many of the markers have been applied successfully to the Bangiales, as well as other lineages previously assigned to the Bangiophyceae sensu lato. All of the primers currently in use with their respective amplification profiles and strategies are provided, which can include full fragment, overlapping fragments and what might best be called "informed overlapping fragments", i.e., a fragment for a marker is amplified and sequenced for a taxon and those sequence data are then used to identify the best primers to amplify the remaining fragment(s) for that marker. We extend this strategy for the more variable markers with sequence from the external PCR primers used to "inform" the selection of internal sequencing primers. This summary will hopefully serve as a useful resource to systematists in the red algal community.

Conditional Replication of a Recombinant Adenovirus Studied Using Neomycin as a Selective Marker

  • Xue, Feng;Qi, Yi-Peng;Joshua, Mallam Nock;Lan, Ping;Dong, Chang-Yuan
    • BMB Reports
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    • v.36 no.3
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    • pp.275-281
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    • 2003
  • An E1B-defective adenovirus, named r2/Ad carrying the neo expression cassette, was constructed by homologous recombination. The construction, selection (using neomycin as a selective marker), and propagation of the recombinant virus was performed in human embryonic kidney 293 cells (HEK 293). An in vitro study demonstrated that this recombinant virus has the ability to replicate in and lyse some p53-deficient human tumor cells such as human glioma tumor cells (U251) and human bladder cells (EJ), but not in some cells with functional p53, such as human adenocarcinoma cells (A549) and human fibroblast cells (MRC-5). Also, based on the cytopathic effect (CPE), it was demonstrated, under identical conditions, that the U251 cells were more sensitive to r2/Ad replication than the EJ cells. In this paper, we report that r2/Ad could be very useful in studying the in vitro selective replication of E1B-defective adenovirus and has great potential in cancer gene therapy.

Temporary Semi-Jailing Technique for Coil Embolization of Wide-Neck Aneurysm with Small Caliber Parent Artery Following Incomplete Clipping

  • Byun, Jun Soo;Kim, Jae Kyun;Lee, Hwa Yeon;Hwang, Sung Nam
    • Journal of Korean Neurosurgical Society
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    • v.53 no.4
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    • pp.241-244
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    • 2013
  • The authors describe the use of a self-expandable stent in a temporary deployment for treatment of a very wide-neck A1 segment of anterior cerebral artery (ACA) aneurysm following incomplete clipping. A 39-year-old hypertensive man presenting with seizure-like movement underwent computed tomography, which showed acute subarachnoid hemorrhage and an A1 segment of ACA aneurysm with superior and inferior projection. He underwent surgical clipping of the aneurysm, but superior and posterior portion of wide-neck aneurysm remained. We decided to treat the remnant aneurysm using an endovascular modality. After selection of the aneurysm, coil packing was performed assisted by the temporary semi-jailing technique. The Enterprise stent (Cordis Neurovascular, Miami, FL, USA) was deployed and recaptured repeatedly for angiography to ensure safety of the small caliber parent artery. Successful semi-deployment and recapture of the stent allowed subtotal coil occlusion of the aneurysm with good anatomic and clinical results. No complications were encountered. The stent could be recaptured up to the point where the proximal end of the stent marker was aligned with distal marker band of the microcatheter, approximately 70% of the stent length. The temporary semi-jailing technique is feasible for wide-neck aneurysm with small caliber parent artery.

D-amino Acid Oxidase (DAO) Gene as a Novel Selection Marker for Plant Transformation (새로운 선발 마커 D-아미노산 산화효소 유전자를 이용한 식물 형질전환)

  • Lim, Sun-Hyung;Woo, Hee-Jong;Lee, Si-Myung;Jin, Yong-Moon;Cho, Hyun-Suk
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.31-36
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    • 2007
  • Though higher plants car not metabolize D-amino acid, many prokaryotes and eukaryotes have the D-amino acid metabolism. Therefore, we transformed tobacco plants with D-amino acid oxidase (DAO), which can metabolize D-amino acid, and confirmed that transgenic tobacco plants might metabolize D-amino acid. Transgenic tobacco plants were survived a high concentration of D-serine, however non-transgenic plants were not grown on D-serine medium. From Southern and Northern blot analysis, transgenic tobacco plants selected on D-serine medium were confirmed by insert and expression of transgene. $T_{1}$ tobacco seeds derived $T_{0}$ tobacco plants selfing were grown on D-serine medium and showed normal phenotype compared to wild tobacco plants. Transgenic tobacco plants displayed the metabolic capability of D-serine. Therefore, we suggested that DAO is useful selectable marker gene for plant transformation.

Application of genomics into rice breeding

  • Ando, Ikuo
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.13-13
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    • 2017
  • By the progress of genome sequencing, infrastructures for marker-assisted breeding (MAB) of rice came to be established. Fine mapping and gene isolation have been conducted using the breeding materials derived from natural variations and artificial mutants. Such genetic analysis by the genome-wide dense markers provided us the knowledge about the many genes controlling important traits. We identified several genes or quantitative trait loci (QTL) for heading date, blast resistance, eating quality, high-temperature stress tolerance, and so on. NILs of each gene controlling heading date contribute to elongate the rice harvest period. Determination of precise gene location of blast resistance gene pi21, allowed us to overcome linkage drag, co-introduction of undesirable eating quality. We could also breed the first practical rice cultivar in Japan with a brown planthopper resistance gene bph11 in the genetic back-ground of an elite cultivar. Discovery of major and minor QTLs for good eating quality allowed us to fine-tune of eating quality according to the rice planting area or usage of rice grain. Many rice cultivars have bred efficiently by MAB for several traits, or by marker-assisted backcross breeding through chromosome segment substitution lines (CSSLs) using genetically diverse accessions. We are also systematically supporting the crop breeding of other sectors by MAB or by providing resources such as CSSLs. It is possible to pyramid many genes for important traits by using MAB, but is still difficult to improve the yielding ability. We are performing a Genomic Selection (GS) for improvement of rice biomass and grain yield. We are also trying to apply the genome editing technology for high yield rice breeding.

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Analysis of Korean japonica rice cultivars using molecular markers associated with blast resistance genes

  • Suh, Jung-Pil;Roh, Jae-Hwan;Cho, Young-Chan;Han, Seong-Sook;Jeon, Yong-Hee;Kang, Kyung-Ho;Kim, Yeon-Gyu
    • Korean Journal of Breeding Science
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    • v.40 no.3
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    • pp.215-222
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    • 2008
  • Fifty-two Korean japonica rice cultivars were analyzed for leaf blast resistance and genotyped with 4 STS and 26 SSR markers flanking the specific chromosome sites linked with blast resistance genes. In our analysis of resistance genes in 52 japonica cultivars using STS markers tightly linked to Pib, Pita, Pi5(t) and Pi9(t), the blast nursery reaction of the cultivars possessing the each four major genes were not identical to that of the differential lines. Eight of the 26 SSR markers were associated with resistant phenotypes against the isolates of blast nursery as well as the specific Korean blast isolates, 90-008 (KI-1113), 03-177 (KJ-105). These markers were linked to Pit, Pish, Pib, Pi5(t), Piz, Pia, Pik, Pi18, Pita and Pi25(t) resistance gene loci. Three of the eight SSR markers, MRG5836, RM224 and RM7102 only showed significantly associated with the phenotypes of blast nursery test for two consecutive years. These three SSR markers also could distinguish between resistant and susceptible japonica cultivars. These results demonstrate the usefulness of marker-assisted selection and genotypic monitoring for blast resistance of rice in blast breeding programs.

Population structure analysis of Yeonsan Ogye using microsatellite markers

  • Cho, Sung Hyun;Lee, Seung-Sook;Manjula, Prabuddha;Kim, Minjun;Lee, Seung Hwan;Lee, Jun Heon;Seo, Dongwon
    • Journal of Animal Science and Technology
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    • v.62 no.6
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    • pp.790-800
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    • 2020
  • The Yeonsan Ogye (YO) chicken is a natural heritage of Korea, characterized by black feathers, skin, bones, eyes, and comb. The purebred of YO population has been reared under the natural mating system with no systematic selection and breeding plan. The purpose of this study was to identify the genetic diversity and find the optimal number of population sub-division using 12 polymorphic microsatellite (MS) markers to construct a pedigree-based breeding plan for the YO population. A total of 509 YO birds were used for this study. Genetic diversity and population structure analysis were conducted based on the MS marker genotype information. The overall average polymorphic information content value and expected heterozygosity of the population were 0.586, and 0.642, respectively. The K-mean cluster analysis based on the genetic distance result confirmed that the current YO population can be divided into three ancestry groups. Individuals in each group were evaluated based on their genetic distance to identify the potential candidates for a future breeding plan. This study concludes that a future breeding plan with known pedigree information of selected founder animals, which holds high genetic diversity, could be the best strategy to ensure the conservation of the Korean YO chicken population.

Single nucleotide polymorphism marker combinations for classifying Yeonsan Ogye chicken using a machine learning approach

  • Eunjin, Cho;Sunghyun, Cho;Minjun, Kim;Thisarani Kalhari, Ediriweera;Dongwon, Seo;Seung-Sook, Lee;Jihye, Cha;Daehyeok, Jin;Young-Kuk, Kim;Jun Heon, Lee
    • Journal of Animal Science and Technology
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    • v.64 no.5
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    • pp.830-841
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    • 2022
  • Genetic analysis has great potential as a tool to differentiate between different species and breeds of livestock. In this study, the optimal combinations of single nucleotide polymorphism (SNP) markers for discriminating the Yeonsan Ogye chicken (Gallus gallus domesticus) breed were identified using high-density 600K SNP array data. In 3,904 individuals from 198 chicken breeds, SNP markers specific to the target population were discovered through a case-control genome-wide association study (GWAS) and filtered out based on the linkage disequilibrium blocks. Significant SNP markers were selected by feature selection applying two machine learning algorithms: Random Forest (RF) and AdaBoost (AB). Using a machine learning approach, the 38 (RF) and 43 (AB) optimal SNP marker combinations for the Yeonsan Ogye chicken population demonstrated 100% accuracy. Hence, the GWAS and machine learning models used in this study can be efficiently utilized to identify the optimal combination of markers for discriminating target populations using multiple SNP markers.

Construction of an Integrated Pepper Map Using RFLP, SSR, CAPS, AFLP, WRKY, rRAMP, and BAC End Sequences

  • Lee, Heung-Ryul;Bae, Ik-Hyun;Park, Soung-Woo;Kim, Hyoun-Joung;Min, Woong-Ki;Han, Jung-Heon;Kim, Ki-Taek;Kim, Byung-Dong
    • Molecules and Cells
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    • v.27 no.1
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    • pp.21-37
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    • 2009
  • Map-based cloning to find genes of interest, marker-assisted selection (MAS), and marker-assisted breeding (MAB) all require good genetic maps with high reproducible markers. For map construction as well as chromosome assignment, development of single copy PCR-based markers and map integration process are necessary. In this study, the 132 markers (57 STS from BAC-end sequences, 13 STS from RFLP, and 62 SSR) were newly developed as single copy type PCR-based markers. They were used together with 1830 markers previously developed in our lab to construct an integrated map with the Joinmap 3.0 program. This integrated map contained 169 SSR, 354 RFLP, 23 STS from BAC-end sequences, 6 STS from RFLP, 152 AFLP, 51 WRKY, and 99 rRAMP markers on 12 chromosomes. The integrated map contained four genetic maps of two interspecific (Capsicum annuum 'TF68' and C. chinense 'Habanero') and two intraspecific (C. annuum 'CM334' and C. annuum 'Chilsungcho') populations of peppers. This constructed integrated map consisted of 805 markers (map distance of 1858 cM) in interspecific populations and 745 markers (map distance of 1892 cM) in intraspecific populations. The used pepper STS were first developed from end sequences of BAC clones from Capsicum annuum 'CM334'. This integrated map will provide useful information for construction of future pepper genetic maps and for assignment of linkage groups to pepper chromosomes.