• Title/Summary/Keyword: SNP marker

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Development of Cleaved Amplified Polymorphic Sequence Markers for the Identification of Lentinula edodes Cultivars Sanmaru 1ho and Chunjang 3ho (표고버섯 품종 산마루1호, 천장3호를 구분할 수 있는 CAPS Marker 개발)

  • Moon, Suyun;Lee, Hwa-Yong;Kim, Myungkil;Ka, Kang-Hyeon;Ko, Han Kyu;Chung, Jong-Wook;Koo, Chang-Duck;Ryu, Hojin
    • The Korean Journal of Mycology
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    • v.45 no.2
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    • pp.114-120
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    • 2017
  • Lentinula edodes is an edible mushroom that is mainly cultivated in Asian countries. Recently, new cultivars of this mushroom have been developed in Korea; variety protection is very important, so the development of efficient molecular markers that can distinguish each variety is required. In this study, we developed cleaved amplified polymorphic sequence (CAPS) markers for the identification of L. edodes cultivars (Sanmaru 1ho and Chunjang 3ho). These markers were developed from whole genomic sequencing data from L. edodes monokaryon strain B17 and resequencing data from 10 dikaryon strains. A single nucleotide polymorphism changed in scaffold 9 POS 1630048 in Sanmaru 1ho($G{\rightarrow}T$), and in scaffold 13 POS 920681 in Chunjang 3ho ($G{\rightarrow}A$). The restriction enzymes TspR I and Xho I distinguished Sanmaru 1ho and Chunjang 3ho, respectively, from other strains. Thus, we developed 2 CAPS markers for the identification of the L. edodes cultivars Sanmaru 1ho and Chunjang 3ho.

An Overview for Molecular Markers in Plants (식물에서 분자 마커의 동향)

  • Huh, Man Kyu
    • Journal of Life Science
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    • v.25 no.7
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    • pp.839-848
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    • 2015
  • A molecular marker is a molecule contained within a sample taken from an organism or other matter. The development of molecular techniques for genetic analysis has led to a great contribution to our knowledge of plant genetics and our understanding of the structure and behavior of various genomes in plants. Recently, functional molecular markers have been developed to detect the presence of major genes from the analysis of pedigreed data in absence of molecular information. DNA markers have developed into many systems based on different polymorphism-detecting techniques or methods such as RFLP, AFLP, RAPD, SSR, SNP, etc. A new class of very useful DNA markers called genic molecular markers utilizing the ever-increasing archives of gene sequence information being accumulated under the EST sequencing projects on a large number of plant species. Functional markers are derived from polymorphic sequences, and are more likely to be involved in phenotypic trait variation. Based on this conceptual framework, the marker systems discussed below are all (gene)-targeted markers, which have the potential to become functional. These markers being part of the cDNA/EST-sequences, are expected to represent the functional component of the genome i.e., gene(s), in contrast to all other random DNA based markers that are developed/generated from the anonymous genomic DNA sequences/domains irrespective of their genic content/information. Especially I sited Poczai et al’ reviews, advances in plant gene-targeted and functional markers. Their reviews may be some useful information to study molecular markers in plants.

An Introduction to Microsatellite Development and Analysis (Microsatellite 개발 및 분석법에 대한 소개)

  • Yun Young-Eun;Yu Jeong-Nam;Lee Byoung-Yoon;Kwak Myounghai
    • Korean Journal of Plant Taxonomy
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    • v.41 no.4
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    • pp.299-314
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    • 2011
  • The choice of molecular markers is the first step when selecting experimental plans in the field of population genetics. The popular molecular markers in population genetic studies are mainly allozyme, RAPD, RFLP, AFLP, microsatellite, SNP and ISSR. Among these, microsatellites are frequently found in nuclear, chloroplast and mitochondrial genome, showing a high level of polymorphism and nuclear microsatellites are codominant. Thus, it is a favorable molecular marker for population structure analyses and genetic diversity studies. Microsatellites are composed of tandem repeated 1~6 base pair nucleotide motifs and can be easily amplified by PCR reactions using locus specific primers. Because microsatellites have low cross-species transferability, however, they are only applicable between phylogenetically close species. In wild plants, the lack of genomic information and the high development cost of the microsatellite obstruct the wider use of microsatellites in plant population genetics research. In this review, we introduce the basis for microsatellite markers, the development process, and analytical methods as well as evolutionary models and their applications. In addition, possible genotyping errors which lead to erroneous conclusions are discussed.

Construction of a Genetic Map using the SSR Markers Derived from "Wonwhang" of Pyrus pyrifolia (배 '원황'(Pyrus pyrifolia) 유전체 해독에 기반한 SSR 마커 개발 및 유전자 지도 작성)

  • Lee, Ji Yun;Seo, Mi-Suk;Won, So Youn;Lim, Kyoung Ah;Shin, Il Sheob;Choi, Dongsu;Kim, Jung Sun
    • Korean Journal of Breeding Science
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    • v.50 no.4
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    • pp.434-441
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    • 2018
  • High-density genetic linkage mapping is critical for undertaking marker-assisted selection and confirming quantitative trait loci, as well as helping to build pseudomolecules of genomes. We constructed a genetic map using 94 $F_1$ populations generated from the interspecific cross between Korean cultivar "Wonwhang" (Pyrus pyrifolia, NCBI BioSample SAMN05196235) and European cultivar "Bartlett" (Pyrus communis). We designed a total of 24,267 SSR markers based on the genome sequences of "Wonwhang" for this. To select the markers that are linked to the traits important in pear breeding programs, SSR-containing genomic sequences were subjected to nucleotide sequence homology searches, which resulted in 510 SSR markers with high similarity to genes encoding proteins with putative functions such as transcription factors, resistance proteins, flowering time, and regulatory genes. Of these, 70 markers showed polymorphisms in parents and segregating populations and were used to construct a genetic linkage map, together with the unpublished 579 SNPs obtained from genotyping by sequencing analysis. The genetic linkage map covered 3,784.2 cM and the average distance between adjacent markers was 5.8 cM. Seventy SSR markers were distributed across 17 chromosomes with more than one locus.

Identification of Novel Single Nucleotide Polymorphisms on ADSL Gene Using Economic Traits in Korean Native Chicken (한국재래닭의 ADSL 유전자 내 단일염기변이를 이용한 경제형질과의 연관성 분석)

  • Lee, J.A.;Jeon, S.A.;Oh, J.D.;Park, K.D.;Choi, K.D.;Jeon, G.J.;Lee, H.K.;Kong, H.S.
    • Korean Journal of Poultry Science
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    • v.36 no.3
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    • pp.207-213
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    • 2009
  • Adenylosuccinate lyase (ADSL) deficiency is a disease of purine metabolism which affects patients both biochemicall and behaviorally. An obstacle of this purine nucleotide cycle(PNC) can be caused brain functional disorder and growth disorder. So ADSL deficiency, which is associated with sever mental retardation, autistic features and energy metabolism. This study was performed to identify SNP on ADSL gene in chicken. The nucleotides were observed as T to C ($7724^{th}$ nucleotide), C to T ($7732^{nd}$ nucleotide), G to T ($10108^{th}$ nucleotide), A to T ($10356^{th}$ nucleotide), G to A($10375^{th}$ nucleotide), A to C ($10402^{nd}$ nucleotide), A to T ($12716^{th}$ nucleotide), T to A ($12717^{th}$ nucleotide), C to T ($15491^{st}$ nucleotide), C to T ($15542^{nd}$ nucleotide) and C to T ($15550^{th}$ nucleotide). The nucleotide substitutions at $15542^{nd}$ and $15550^{th}$ (GeneBank accession no. AY665559) were found as missense mutation (alanine$\rightarrow$valine, proline$\rightarrow$serine, respectively). This study will be useful for farther researches for identifying association between these SNPs and energy metabolism in chicken. The C15550T SNP showed three genotypes, CC, CT, TT by digestion with the genotype TT had significantly faster the first lay day (150.0) than CT (162.0, P<0.05) and genotype TT (150.0, P<0.05) had significantly higher the egg production rate than CT (172.4, P<0.05). According to result of this study, a C15550T was found to have a significantly effect first lay day and mean egg production. It will be possible to use SNP marker on selecting chicken to improve important economic traits, which is the first lay day and mean egg production.

EST Profiling for Seed-hair Characteristic and Development of EST-SSR and SNP Markers in Carrot (당근 종모 형질 관련 EST profiling과 이를 이용한 EST-SSR 및 SNP 마커 개발)

  • Oh, Gyu-Dong;Hwang, Eun-Mi;Shim, Eun-Jo;Jeon, Sang-Jin;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.28 no.6
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    • pp.1025-1038
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    • 2010
  • Carrot ($Daucus$ $carota$ L. var. $sativa$) is one of the most widely used crops in the world. Moreover it is an important crop because of its high content of ${\beta}$-carotene, well-known as the precursor of vitamin A carotenoid. However, seed-hair which is generated in epidermal cell of seeds inhibits absorption and germination. For that reason, carrot seeds are commercialized after mechanical hair removal process. To overcome such cumbersome weaknesses, new breeding program for developing hairless-seed carrot cultivar has been needed. Therefore, in this study, cDNA libraries from seeds of short-hair seed phenotype CT-ATR615 OP 666-13line and hairy seed CT-ATR615 OP-CK1-9 line were constructed and expression patterns related to generation of seed-hair were analyzed by comparison of EST sequences. Differential EST sequence results between two lines were classified into FunCat functional categories based on the results of BlastX search. Higher expression quantities belonging to metabolic category were shown on short-hair seed line than hairy-seed one. Differential expression quantities between those two lines in the protein folding and stabilization, subcellular localization categories were supposed to contribute variously on the generation of seed-hair. We confirmed 50 and 59 SSR sites, and 2 SNP sites by analyzing EST sequences in two lines; thereafter, we designed SNP and SSR primer sets from these EST sequence information as a molecular marker. These markers are thought to be used in research of molecular markers for classification of carrot family and related to various traits, as well as seed-hair characteristic.

Characterization of the Lsi1 Homologs in Cucurbita moschata and C. ficifolia for Breeding of Stock Cultivars Used for Bloomless Cucumber Production

  • Jung, Jaemin;Kim, Joonyup;Jin, Bingkui;Choi, Youngmi;Hong, Chang Oh;Lee, Hyun Ho;Choi, Youngwhan;Kang, Jumsoon;Park, Younghoon
    • Horticultural Science & Technology
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    • v.35 no.3
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    • pp.333-343
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    • 2017
  • Bloomless cucumber fruits are commercially produced by grafting onto the pumpkin stocks (Cucurbita moschata) to restricted silicon ($SiO_2$) absorption. Inhibition of silicon absorption in bloomless stocks is conferred by a mutant allele of the CmLsi1 homologous to Lsi1 in rice. In this study, we characterized the Lsi1 homologs in pumpkin (C. moschata) and its cold-tolerant wild relative C. ficifolia ('Heukjong') in order to develop a DNA marker for selecting a bloomless trait and to establish the molecular basis for breeding bloomless stock cultivars of C. ficifolia. A Cleaved amplified polymorphic sequence (CAPS) marker (CM1-CAPS) was designed based on a non-sysnonymous single nucleotide polymorphism (SNP, C>T) of the CmLsi1 mutant-type allele, and its applicability for Marker-assisted selection (MAS) was confirmed by evaluating three bloom and five bloomless pumpkin stock cultivars. Quantitative RT-PCR of the CmLsi1 for these stock cultivers implied that expression level of the CmLsi1 gene does not appear to be associated with the bloom/bloomless trait and may differ depending on plant species and tissues. A full length cDNA of the Lsi1 homolog [named CfLsi1($B^+$)] of 'Heukjong' (C. ficifolia), was cloned and sequence comparison between CmLsi1($B^+$) and CfLsi1($B^+$) revealed that there exists total 24 SNPs, of which three were non-synonymous. Phylogenetic analysis of CfLsi1($B^+$) and Lsi1 homologs further revealed that CfLsi1($B^+$) is closesly related to Nodulin 26-like intrinsic proteins (NIPs) and most similar to CpNIP1 of C. pepo than C. moschata.

Characterization of Phosphoinositide-3-kinase, Class 3 (PIK3C3) Gene and Association Tests with Quantitative Traits in Pigs

  • Kim, J.H.;Choi, B.H.;Lim, H.T.;Park, E.W.;Lee, S.H.;Seo, B.Y.;Cho, I.C.;Lee, J.G.;Oh, S.J.;Jeon, J.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.12
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    • pp.1701-1707
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    • 2005
  • This study deals with the characterization of porcine PIK3C3 and association tests with quantitative traits. PIK3C3 belongs to the class 3 PI3Ks that participate in the regulation of hepatic glucose output, glycogen synthase, and antilipolysis in typical insulin target cells such as those in the such as liver, muscle system, and fat. On the analysis of full-length mRNA sequence, the length of the PIK3C3 CDS was recorded as 2,664 bps. As well, nucleotide and amino acid identities between human and pig subjects were 92% and 99%, respectively. Five SNPs were detected over 5 exons. We performed genotyping by using a SNP C2604T on exon24 for 145 F$_2$ animals (from a cross between Korean native boars and Landrace sows) by PCR-RFLP analysis with Hpy8I used to investigate the relationship between growth and fat depot traits. In the total association analysis, which doesn' consider transmission disequilibrium, the SNP showed a significant effect (p<0.05) on body weight and carcass fat at 30 weeks of age as well as a highly significant effect (p<0.01) on back fat. In an additional sib-pair analysis, C allele still showed positive and significant effects (p<0.05) on back fat thickness and carcass fat. Moreover, the effects of C allele on the means of within-family components for carcass fat and back fat were estimated as 2.76 kg and 5.07 mm, respectively. As a result, the SNP of porcine PIK3C3 discovered in this study could be utilized as a possible genetic marker for the selection of pigs that possess low levels of back fat and carcass fat at the slaughter weight.

Association Study Analysis of Cluster-of-Differentiation Antigen 9 (CD9) Gene Polymorphism (g.358A>T) for Duroc Boar Post-thawed Semen Motility and Kinematic Characteristics

  • Cho, Eun-Seok;Sa, Soo-Jin;Kim, Ki-Hyun;Lee, Mi-Jin;Ko, Jun-Ho;Kim, Young-Ju;Seol, Kuk-Hwan;Hong, Joon-ki;Kim, Kwang-Sik;Kim, Yong-Min;Woo, Jae-Seok
    • Journal of Embryo Transfer
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    • v.30 no.2
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    • pp.109-114
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    • 2015
  • Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Various researches have been trying to improve the quality of semen post-thawed in boar. Recently, polymorphism (g.358A>T) of cluster-of-differentiation antigen 9 (CD9) gene reported to be significant association with MOT. Also, CD9 gene was expressed in the male germ line stem cells is crucial for sperm-egg fusion, and was therefore selected as candidate gene for boar semen. This study was conducted to evaluate the pig SNP (g.358A>T) of CD9 gene as a positional controlling for semen parameters of post-thawed boar semen. To results, the g.358A>T SNP of the CD9 gene was significantly associated with the traits such as MOT, curve linear velocity, straight line velocity, average path velocity and amplitude of lateral head displacement. Particularly, the g.358A>T SNP significantly has the highest association with MOT and animals with AA genotype (p<0.001). Therefore, we suggest that the g.358A>T in the intron 6 region of the porcine CD9 may be used as a molecular marker for Duroc boar Post-thawed semen quality, although its functional effect was not defined yet.

Single Nucleotide Polymorphism in the Coding Region of Bovine Chemerin Gene and Their Associations with Carcass Traits in Japanese Black Cattle