• Title/Summary/Keyword: ITS marker

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Characterization of the porcine Nanog 5'-flanking region

  • Memon, Azra;Song, Ki-Duk;Lee, Woon Kyu
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.3
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    • pp.449-456
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    • 2018
  • Objective: Nanog, a homeodomain protein, has been investigated in humans and mice using embryonic stem cells (ESCs). Because of the limited availability of ESCs, few studies have reported the function and role of Nanog in porcine ESCs. Therefore, in this study, we investigated the location of the porcine Nanog chromosome and its basal promoter activity, which might have potential applications in development of ESCs specific marker as well as understanding its operating systems in the porcine. Methods: To characterize the porcine Nanog promoter, the 5'-flanking region of Nanog was isolated from cells of mini-pig ears. BLAST database search showed that there are two porcine Nanog genomic loci, chromosome 1 and 5, both of which contain an exon with a start codon. Deletion mutants from the 5'-flanking region of both loci were measured using the Dual-Luciferase Reporter Assay System, and a fluorescence marker, green fluorescence protein. Results: Promoter activity was detected in the sequences of chromosome 5, but not in those of chromosome 1. We identified the sequences from -99 to +194 that possessed promoter activity and contained transcription factor binding sites from deletion fragment analysis. Among the transcription factor binding sites, a Sp1 was found to play a crucial role in basal promoter activity, and point mutation of this site abolished its activity, confirming its role in promoter activity. Furthermore, gel shift analysis and chromatin immunoprecipitation analysis confirmed that Sp1 transcription factor binds to the Sp1 binding site in the porcine Nanog promoter. Taken together, these results show that Sp1 transcription factor is an essential element for porcine Nanog basal activity the same as in human and mouse. Conclusion: We showed that the porcine Nanog gene is located on porcine chromosome 5 and its basal transcriptional activity is controlled by Sp1 transcription factor.

Expression of DNA Methylation Marker of Paired-Like Homeodomain Transcription Factor 2 and Growth Receptors in Invasive Ductal Carcinoma of the Breast

  • Rahman, Wan Faiziah Wan Abdul;Fauzi, Mohd Hashairi;Jaafar, Hasnan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.19
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    • pp.8441-8445
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    • 2014
  • Background: Paired-like homeodomain transcription factor 2 (PITX2) is another new marker in breast carcinoma since hypermethylation at P2 promoter of this gene was noted to be associated with poor prognosis. We investigated the expression of PITX2 protein using immunohistochemistry in invasive ductal carcinoma and its association with the established growth receptors such as estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth receptor 2 (HER2). Methods: We conducted a cross sectional study using 100 samples of archived formalin-fixed paraffin embedded tissue blocks of invasive ductal carcinoma and stained them with immunohistochemistry for PITX2, ER, PR and HER2. All HER2 with scoring of 2+ were confirmed with chromogenic in-situ hybridization (CISH). Results: PITX2 protein was expressed in 53% of invasive ductal carcinoma and lack of PITX2 expression in 47%. Univariate analysis revealed a significant association between PITX2 expression with PR (p=0.001), ER (p=0.006), gland formation (p=0.044) and marginal association with molecular subtypes of breast carcinoma (p=0.051). Combined ER and PR expression with PITX2 was also significantly associated (p=0.003) especially in double positive cases. Multivariate analysis showed the most significant association between PITX2 and PR (RR 4.105, 95% CI 1.765-9.547, p=0.001). Conclusion: PITX2 is another potential prognostic marker in breast carcinoma adding significant information to established prognostic factors of ER and PR. The expression of PITX2 together with PR may carry a very good prognosis.

RAPD marker variations between and within the species of Korean Suaeda (한국산 나문재속의 종내·종간 RAPD marker 변이)

  • Shim, Hyun-Bo;Choi, Byoung-Hee
    • Korean Journal of Plant Taxonomy
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    • v.34 no.1
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    • pp.63-74
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    • 2004
  • The genus Suaeda is one of the most popular plants on salt marsh areas in Korean Peninsular. The entities that comprise taxa in Korea exhibit widely overlapping ranges in all morphological attributes. Ramdom amplified polimorphic DNA (RAPD) markers were used to clarify taxonomic delimitation among the Korean taxa of Suaeda and to analyse genetic variations among the populations of S. japoniro in western and southern coastal regions. Six decamer primers amplified a total of 65 scorable bands, of which 61 were polymorphic. In all primers investigated, S. glauro of sect. Schanginia is the most distinctive species, compared with others of sect. Heterosperma. S. japonica, S. maritima, and S. malacosperma, which have been hardly distinguished each other by external morphology, were readily recognized by its specific DNA bands. The characteristic RAPD markers were identified local populations of S. japonica, but this feature was not revealed within population.

Genetic diversity, structure analysis and relationship in soybean mutants as revealed by TRAP marker

  • Kim, Dong-Gun;Lyu, Jae-Il;Lee, Min-Kyu;Kim, Jung Min;Hong, Min Jeong;Kim, Jin-Baek;Bae, Chang-Hyu;Kwon, Soon-Jae
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.43-43
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    • 2018
  • Mutation breeding by radiation is useful for improving various crop species. Up to now, a total of 170 soybean mutant varieties have been released in the world, which is the second most registered varieties after rice. Despite the economic importance of soybean, there have been no TRAP marker system studies on genetic relationships between/among mutant lines. To develop a strategy of Mutant Diversity Pool (MDP) conservation, a study on the genetic diversity of 210 soybean mutant lines (8 cultivars and 202 mutants) was performed through a TRAP analysis. Sixteen primer combinations amplified a total of 551 fragments. The highest (84.00%) and lowest (32.35%) polymorphism levels were obtained with primers MIR157B + Ga5 and B14G14B + Ga3, respectively. The mean PIC values 0.15 varied among the primer combination ranging from 0.07 in B14G14B + Sal2 to 0.23 in MIR157B + Sa4. Phylogenetic, principal component analysis (PCA) and structure analysis indicated that the 210 lines belong to four groups based on the 16 combination TRAP markers. AMOVA showed 21.0% and 79.0% variations among and within the population, respectively. Overall, the genetic similarity of each cultivar and its mutants were higher than within other mutant populations. Our results suggest that the TRAP marker system may be useful for assessing the genetic diversity among soybean mutants and help to improve our knowledge of soybean mutation breeding.

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Age-Related Changes of Adult Neural Stem Cells in the MouseHippocampal Dentate Gyrus

  • Jung, Ji-Yeon;Byun, Kang-Ok;Jeong, Yeon-Jin;Kim, Won-Jae
    • International Journal of Oral Biology
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    • v.33 no.2
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    • pp.59-64
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    • 2008
  • This study was designed to investigate the changes in the properties of the neuronal setm cells or progenitor cells associated with age-related decline in neurogenesis of the hippocampal dentate gyrus (DG). Active whole cells cycle marker Ki67 (a marker of whole cell cycle)-positive and S phase marker bromodeoxyuridine (BrdU)-positive. Neural stem cells gradually were reduced in the hippocampal subgranular zone (SGZ) in an age-dependant manner after birth (from P1 month to P1 year). The ratio of BrdUpositivecells/Ki67-positive cells was gradually enhanced in an age-dependent manner. The ratio of Ki67-positive cells/accu-mulating BrdU-positive cells at 3 hrs after BrdU injection was injected once a day for consecutive 5 days gradually decreased during ageing. TUNEL- and caspase 3 (apoptotic terminal caspase)-positive cells gradually decreased in the dentate SGZ during ageing and immunohistochemical findings of glial fibrillary acid protein (GFAP) were not changed during ageing. NeuN, a marker of mature neural cells, and BrdU-double positive cells gradually decreased in an age-dependent manner but differentiating ratio and survival rate of cells were not changed at 4 wks after BrdU injection once a day for consecutive 5 days. The number of BrdU-positive cells migrated from the hippocampal SGZ into granular layer and its migration speed was gradually declined during ageing. These results suggest that the adult neurogenesis in the mouse hippocampal DG gradually decrease through reducing proliferation of neural stem cells accompanying with cells cycle change and reduced cells migration rather than changes of differentiation.

Identification of a Novel SNP Associated with Meat Quality in C/EBP${\alpha}$ Gene of Korean Cattle

  • Shin, S.C.;Kang, M.J.;Chung, E.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.4
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    • pp.466-470
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    • 2007
  • CCAAT/enhancer binding protein ${\alpha}$($C/EBP{\alpha}$) plays an important role in lipid deposition and adipocyte differentiation. In order to find genetic markers to improve the meat quality of Korean cattle, the bovine $C/EBP{\alpha}$ gene was chosen as a candidate gene to investigate its association with carcass and meat quality traits in Korean cattle. A single nucleotide polymorphism (SNP) was identified at position 271 (A/C substitution) of coding region in the $C/EBP{\alpha}$ gene. A PCR-RFLP procedure with restriction enzyme SmaI was developed for determining the marker genotypes. The frequencies of alleles C and A and were 0.374 and 0.626, respectively. The genotype frequencies for CC, AC and AA were 12.9, 49.0 and 38.1%, respectively, in Korean cattle population. The frequencies of genotype were in agreement with Hardy-Weinberg equilibrium. Association analysis indicated that the gene-specific SNP marker of $C/EBP{\alpha}$ showed a significant association with marbling score (p<0.05). The animals with AA genotype had higher marbling score than those with the AC or CC genotype. Although further studies are needed to validate our results, the $C/EBP{\alpha}$ gene could be useful as a genetic marker for carcass and meat quality traits in Korean cattle.

Predicting the Accuracy of Breeding Values Using High Density Genome Scans

  • Lee, Deuk-Hwan;Vasco, Daniel A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.2
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    • pp.162-172
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    • 2011
  • In this paper, simulation was used to determine accuracies of genomic breeding values for polygenic traits associated with many thousands of markers obtained from high density genome scans. The statistical approach was based upon stochastically simulating a pedigree with a specified base population and a specified set of population parameters including the effective and noneffective marker distances and generation time. For this population, marker and quantitative trait locus (QTL) genotypes were generated using either a single linkage group or multiple linkage group model. Single nucleotide polymorphism (SNP) was simulated for an entire bovine genome (except for the sex chromosome, n = 29) including linkage and recombination. Individuals drawn from the simulated population with specified marker and QTL genotypes were randomly mated to establish appropriate levels of linkage disequilibrium for ten generations. Phenotype and genomic SNP data sets were obtained from individuals starting after two generations. Genetic prediction was accomplished by statistically modeling the genomic relationship matrix and standard BLUP methods. The effect of the number of linkage groups was also investigated to determine its influence on the accuracy of breeding values for genomic selection. When using high density scan data (0.08 cM marker distance), accuracies of breeding values on juveniles were obtained of 0.60 and 0.82, for a low heritable trait (0.10) and high heritable trait (0.50), respectively, in the single linkage group model. Estimates of 0.38 and 0.60 were obtained for the same cases in the multiple linkage group models. Unexpectedly, use of BLUP regression methods across many chromosomes was found to give rise to reduced accuracy in breeding value determination. The reasons for this remain a target for further research, but the role of Mendelian sampling may play a fundamental role in producing this effect.

Contact Sensitivity to Dinitrochlorobenzene as a Marker Trait in the Indirect Selection for Body Mange and Coccidiosis Resistance in Broiler Rabbits

  • Nandakumar, P.;Thomas, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.2
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    • pp.165-168
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    • 1999
  • To determine the effects of genetic and environmental influences on cell mediated immune (CMI) responses in broiler rabbits, contact sensitivity to 2,4-Dinitrochlorobenzene (DNCB) was assessed in three temperate broiler breeds of rabbits, namely Soviet Chinchilla, New Zealand White and Grey Giant. The feasibility of using the contact sensitivity to DNCB as a marker trait in selection for disease resistance was examined. There were highly significant differences between breeds (p<0.01) in initial skin thickness and contact sensitivities to DNCB at 24, 48 and 72 hours. Initial skin thickness was greatest in the Soviet Chinchilla breed (mean 2.2484 mm), and was significantly greater (p<0.01) in males (2.4963 mm) than in females (1.7846 mm) (p<0.01). Highest contact sensitivity to DNCB was in the New Zealand White breed with mean increase in skin thickness of 1.1884, 0.9072 and 0.5879 mm at 24, 48 and 72 hours post challenge respectively. Delayed type hypersensitivity (DTH) reaction to DNCB at 24 hours post challenge had a highly significant association (p<0.01) with the incidence of body mange in rabbits. The results indicated a lowered contact sensitivity to DNCB at 24 hours post challenge was associated significantly (p<0.01) with an increase in incidence and severity of body mange, suggesting its potential value as a marker. The correlation s among contact sensitivities at 24, 48 and 72 hours were positive and highly significant (p<0.01); correlations between initial skin thickness and contact sensitivities were negative and highly significant (p<0.01). Another notable significant correlation was between body weight and delayed type hypersensitivity at 24 hours indicating that an enhanced CMI might be associated with better growth rate and general wellbeing.

Implementation of Infant Learning Content using Augmented Reality (증강현실을 이용한 유아용 학습 콘텐츠의 구현)

  • Lee, Jong-Hyeok;Cho, Hyun-Wook
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.15 no.1
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    • pp.257-263
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    • 2011
  • Recently as AR(Augmented Reality) is focus of attention, AR is applied to various fields and is expected its valuable use. In this paper, we implemented the system based on Goblin XNA which supports high resolution model file and higher AR. We confirmed the relation of model output among the number of marker, the location and changes of camera distance. And we produced the infantile studying contents using AR and embodied. In implemented contents, we showed the familiar character to infants on each page marker. As the result of it, we can raise their concentration and at a time studying supporters can use the contents easily as well. Also we put 3 marker on each page of contents to recognize it smoothly in case one part of it is hidden by any obstacle. Finally we maximized the learning effect such as presence and immersion in studying through reinforcing 3D models according to the every situation.

Molecular Authentication and Phylogenetic Analysis of Plant Species for Breeae and Cirsii Herba based on DNA barcodes (DNA 바코드 분석을 통한 소계(小薊) 및 대계(大薊) 기원식물 감별과 종간 유연관계 분석)

  • Moon, Byeong Cheol;Lee, Young Mi;Ji, Yunui;Choi, Goya;Chun, Jin Mi;Kim, Ho Kyoung
    • The Korea Journal of Herbology
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    • v.28 no.3
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    • pp.75-84
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    • 2013
  • Objectives : The origin of Breeae Herba (So-gye) and Cirsii Herba (Dae-gye) is differently prescribed in Korean and Chinese modern pharmacopoeia. Since the similar morphological characteristics and chaotic plant names, moreover, the aerial part of Carduus crispus have been used as the Cirsii Herba. To develop a reliable method for correct identification of these herbal medicines and to evaluate the genetic relationship of these closely related plant species, we analyzed sequences of DNA barcode regions. Methods : Thirty-one samples of 6 medicinal plants (B. segeta, B. setosa, C. japonicum var. maackii, C. setidens, C. chanroenicum, and C. crispus) were collected from different habitate and nucleotide sequences of DNA barcode regions (rDNA-ITS, matK, and rbcL) were analyzed after amplification using appropriate primers reported in previous studies. The nucleotides of species-specific authentic marker and phylogenetic relations were estimated based on the entire sequences of DNA barcodes by the analysis of ClastalW and UPGMA, respectively. Results : In comparative analysis of DNA barcode sequences, we obtained specific nucleotides to discriminate the medicinal plant of Breeae/Cirsii Herba in species level and evaluated the phylogenetic relationship of these species. Futhermore, we identified distinct marker nucleotides enough to authenticate respective species. These sequence differences at corresponding positions were avaliable genetic markers to determine the botanical origins of Breeae Herbal as well as Cirsii Herba. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by providing of definitive information that can identify each plant species and distinguish from unauthentic adulterants and substitutes.