• Title/Summary/Keyword: sequence repeat

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Evaluation of QTL Related SSR Marker Universality in Korean Rice Breeding Populations

  • Song, Moon-Tae;Lee, Jeom-Ho;Lee, Sang-Bok;Ku, Ja-Hwan;Cho, Youn-Sang;Song, Myung-Hee;Park, Sung-Ho;Hwang, Hung-Goo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.1
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    • pp.56-64
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    • 2003
  • If a quantitative trait loci (QTL) marker identified in a population is applicable to different populations (marker universality), this will not only reduce the labor and cost in marker assisted selection (MAS), but accelerate the application of molecular markers to real breeding programs. Present study aims to evaluate the defined QTL related markers from a population to a different breeding population for the MAS. Four rice breeding populations were subjected to seventy-five simple sequence repeat (SSR) markers which were already identified for their polymorphism information content (PIC) in the parents of the crossings. Among them, eight markers were evaluated for their correlation between presence of marker alleles and phenotypic expression in breeding populations. A reasonable level of polymorphism for the mapped markers originated from any sources of rice accessions was observed between crosses of any sources (marker repeatability). However, correlation between presence of markers and expression of the traits in rice breeding populations was not significant except for minor portion of traits and markers examined (failure of marker universality). In the present study, various strategies were discussed to develop new markers with universality of breeding application.

Development of Simple Sequence Repeat Markers from Adenophora triphylla var. japonica (Regel) H. Hara using Next Generation Sequencing (차세대염기서열분석법을 이용한 잔대의 SSR 마커 개발)

  • Park, Ki Chan;Kim, Young Guk;Hwangbo, Kyeong;Gil, Jinsu;Chung, Hee;Park, Sin Gi;Hong, Chang Pyo;Lee, Yi
    • Korean Journal of Medicinal Crop Science
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    • v.25 no.6
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    • pp.411-417
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    • 2017
  • Background: Adenophora triphylla var. japonica (Regel) H. Hara shows vegetative growth with radical leaves during the first year and shows reproductive growth with cauline leaves and bolting during the second year. In addition, the shape of the plant varies within the same species. For this reason, there are limitations to classifying the species by visual examination. However, there is not sufficient genetic information or molecular tools to analyze the genetic diversity of the plant. Methods and Results: Approximately 34.59 Gbp of raw data containing 342,487,502 reads was obtained from next generation sequencing (NGS) and these reads were assembled into 357,211 scaffolds. A total of 84,106 simple sequence repeat (SSR) regions were identified and 14,133 primer sets were designed. From the designed primer sets, 95 were randomly selected and were applied to the genomic DNA which was extracted from five plants and pooled. Thirty-nine primer sets showing more than two bands were finally selected as SSR markers, and were used for the genetic relationship analysis. Conclusions: The 39 novel SSR markers developed in this study could be used for the genetic diversity analysis, variety identification, new variety development and molecular breeding of A. triphylla.

Analysis of the Dual Promoters and the $H_2O$$_2$-responsive Element of the cats Gene Encoding Catalase A in Streptomyces coelicolor

  • Cho, You-Hee;Hahn, Ji-Sook;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.38 no.4
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    • pp.239-244
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    • 2000
  • The cats gene encodes the major catalase in Sreptomyces coelicolor, whose production increases upon H$_2$O$_2$treatment. Besides the previously identified primary promoter (catApl), a minor promoter (catAp2) was newly assigned by S1 nuclease mapping. The catAp2 transcript was observed transiently upon entry into the stationary phase in liquid culture and upon differentiation on solid plates, whereas the level of catApl transcription did not chance significantly during this growth transition. ThecatApl promoter was transcribed by the major vegetative RNA polymerase holoenzyme containing $\sigma$$\^$HrdB/, whereas the catAp2 was transcribed in vitro by the holoenzyme containing $\sigma$$\^$R/ that is activated under oxidative conditions. The cia-element regulating the H$_2$O$_2$-inducibility of catApl was identified within the 23 bp inverted repeat sequence located between -65 and -43 of the catApl promoter. We roamed this sequence HRE (H$_2$O$_2$-responsive Element). The distal half of the inverted repeat was more crucial for H$_2$O$_2$-dependent induction of the catApl transcript than the proximal half. HRE most likely serves as a binding site for the H$_2$O$_2$-responsive repressor CatR.

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Development of Polymorphic Simple Sequence Repeat Markers using High-Throughput Sequencing in Button Mushroom (Agaricus bisporus)

  • Lee, Hwa-Yong;Raveendar, Sebastin;An, Hyejin;Oh, Youn-Lee;Jang, Kab-Yeul;Kong, Won-Sik;Ryu, Hojin;So, Yoon-Sup;Chung, Jong-Wook
    • Mycobiology
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    • v.46 no.4
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    • pp.421-428
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    • 2018
  • The white button mushroom (Agaricus bisporus) is one of the most widely cultivated species of edible mushroom. Despite its economic importance, relatively little is known about the genetic diversity of this species. Illumina paired-end sequencing produced 43,871,558 clean reads and 69,174 contigs were generated from five offspring. These contigs were subsequently assembled into 57,594 unigenes. The unigenes were annotated with reference genome in which 6,559 unigenes were associated with clusters, indicating orthologous genes. Gene ontology classification assigned many unigenes. Based on genome data of the five offspring, 44 polymorphic simple sequence repeat (SSR) markers were developed. The major allele frequency ranged from 0.42 to 0.92. The number of genotypes and the number of alleles ranged from 1 to 4, and from 2 to 4, respectively. The observed heterozygosity and the expected heterozygosity ranged from 0.00 to 1.00, and from 0.15 to 0.64, respectively. The polymorphic information content value ranged from 0.14 to 0.57. The genetic distances and UPGMA clustering discriminated offspring strains. The SSR markers developed in this study can be applied in polymorphism analyses of button mushroom and for cultivar discrimination.

Analysis of Molecular Variance and Population Structure of Sesame (Sesamum indicum L.) Genotypes Using Simple Sequence Repeat Markers

  • Asekova, Sovetgul;Kulkarni, Krishnanand P.;Oh, Ki Won;Lee, Myung-Hee;Oh, Eunyoung;Kim, Jung-In;Yeo, Un-Sang;Pae, Suk-Bok;Ha, Tae Joung;Kim, Sung Up
    • Plant Breeding and Biotechnology
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    • v.6 no.4
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    • pp.321-336
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    • 2018
  • Sesame (Sesamum indicum L.) is an important oilseed crop grown in tropical and subtropical areas. The objective of this study was to investigate the genetic relationships among 129 sesame landraces and cultivars using simple sequence repeat (SSR) markers. Out of 70 SSRs, 23 were found to be informative and produced 157 alleles. The number of alleles per locus ranged from 3 - 14, whereas polymorphic information content ranged from 0.33 - 0.86. A distance-based phylogenetic analysis revealed two major and six minor clusters. The population structure analysis using a Bayesian model-based program in STRUCTURE 2.3.4 divided 129 sesame accessions into three major populations (K = 3). Based on pairwise comparison estimates, Pop1 was observed to be genetically close to Pop2 with $F_{ST}$ value of 0.15, while Pop2 and Pop3 were genetically closest with $F_{ST}$ value of 0.08. Analysis of molecular variance revealed a high percentage of variability among individuals within populations (85.84%) than among the populations (14.16%). Similarly, a high variance was observed among the individuals within the country of origins (90.45%) than between the countries of origins. The grouping of genotypes in clusters was not related to their geographic origin indicating considerable gene flow among sesame genotypes across the selected geographic regions. The SSR markers used in the present study were able to distinguish closely linked sesame genotypes, thereby showing their usefulness in assessing the potentially important source of genetic variation. These markers can be used for future sesame varietal classification, conservation, and other breeding purposes.

Genetic diversity and population structure of European button mushroom (Agaricus bisporus) using SSR markers (SSR 마커를 이용한 유럽 양송이 자원의 유전적 다양성 및 집단구조분석)

  • Shin, Hye-Ran;An, Hyejin;Bang, Jun Hyoung;Kim, Jun Je;Han, Seahee;Lee, Hwa-Yong;Chung, Jong-Wook
    • Journal of Mushroom
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    • v.18 no.4
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    • pp.323-330
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    • 2020
  • Agaricus bisporus is an important edible mushroom that is used as a functional food. In this study, European A. bisporus strains were analyzed for genetic diversity, population structure, and genetic differentiation using simple sequence repeat (SSR) markers. European A. bisporus strains were divided into four groups by distance-based analysis and two subpopulations by model-based analysis. The SSR markers used in this study did not group European A. bisporus strains by geographical region or pileus color. Genetic diversity was high in Group 4 based on distance-based analysis and Pop. 2 based on model-based analysis. A. bisporus strains showed very low genetic differentiation. The results of this study can be used for breeding A. bisporus in the future.

In vitro micropropagation of M.26 (Malus pumila Mill) apple rootstock and assessment of the genetic diversity of proliferated plantlets using simple sequence repeat markers (사과 대목 M.26 (Malus pumila Mill)의 기내 대량번식 및 simple sequence repeat 마커를 이용한 증식된 식물체의 유전적 다양성 평가)

  • Cho, Kang Hee;Han, Bong Hee;Han, Jeom Hwa;Park, Seo Jun;Kim, Se Hee;Lee, Han Chan;Kim, Mi Young;Kim, Myung-Su
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.382-391
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    • 2018
  • The objective of this study was to determine the most effective medium condition of shoot proliferation and root formation for the efficient in vitro micropropagation of M.26 (Malus pumila Mill). Simple sequence repeat (SSR) markers were used to analyze the genetic diversity of micro-propagated and greenhouse grown M.26. Shoot proliferation was carried out in MS (Murashige and Skoog) containing benzyladenin (BA, $0.5{\sim}5.0mg{\cdot}L^{-1}$) and thidiazuron (TDZ, $0.01{\sim}0.1mg{\cdot}L^{-1}$). The highest number of shoots (10.67 shoots per explant) was induced by adding BA at a concentration $1.0mg{\cdot}L^{-1}$. TDZ treatments caused higher hyperhydricity rate in cultured explants than in BA treatments. There was no significant effect of both BA and auxin on shoot proliferation, and the optimum proliferation medium for M.26 was MS medium containing $1.0mg{\cdot}L^{-1}$ BA. To find a suitable medium composition for shoot rooting, we tested different concentrations indole-3-butyric acid (IBA) and ${\alpha}$-naphthaleneacetic acid ($0.5{\sim}5.0mg{\cdot}L^{-1}$), MS medium (1/4-1), sucrose ($0{\sim}30g{\cdot}L^{-1}$). The shoots showed good rooting on half-strength MS medium containing $1.0mg{\cdot}L^{-1}$ IBA and $15-20g{\cdot}L^{-1}$ sucrose. The rooting rate (100%), number of roots (10.45 ~ 13.60 roots per explant), root length (7.41 ~ 8.33 cm), and shoot length (4.93 ~ 5.38 cm) were good on this medium. Fifteen SSR primers were detected in a total of 30 alleles in 20 micro-propagated plantlets, all SSR profiles from micro-propagated plantlets were monomorphic and similar to greenhouse grown control plantlet M.26 plant. The results indicated that M.26 micro-propagated plantlets were genetically stable.

Identification of Genetic Markers for Korean Native Cattle (Hanwoo) by RAPD Analysis

  • Yeo Jung Sou;Lee Ji Sun;Lee Chang Hee;Jung Young Ja;Nam Doo Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.1
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    • pp.23-26
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    • 2000
  • In order to develop the specific genetic marker for Korean native cattle (Hanwoo), randomly amplified polymorphic DNA (RAPD) analysis of 6 different cattle breeds was attempted by using 38 decamer primers. In comparison of RAPD patterns, two distinctive DNA bands specific for Hanwoo were detected. One was 296 bp of DNA fragment found to be specific only for female Hanwoo when primer GTCCACACGG was employed. In individual analysis of this RAPD marker was observed only in female individuals with the possibility of $85.3\%$. The other was 521 bp of RAPD marker amplified using TCGGCGATAG and AGCCAGCGAA primers, which showed $83.0\%$ of genetic frequency in 85 male and 68 female individuals tested. Nucleotide sequencing of these genetic markers revealed that 296 bp marker has a short micro satellite-like sequence, ACCACCACAC, and a tandem repeat sequence of microsatellite GAAAAATG in the determined sequence. Two distinctive tandem repeats of microsatellite sequences, MC and GAAGA, were also appeared in 521 bp DNA marker. In BLAST search, any gene having high homology with these markers was not found.

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Development and evaluation of semi-nested PCR for detection of the variable lipoprotein haemagglutinin (vlhA) gene of Mycoplasma Synoviae in chicken

  • Pohuang, Tawatchai;Phuektes, Patchara;Junnu, Sucheeva
    • Korean Journal of Veterinary Research
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    • v.60 no.3
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    • pp.109-116
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    • 2020
  • This study aimed to develop a semi-nested polymerase chain reaction assay for the direct detection of Mycoplasma synoviae (M. synoviae) from clinical samples using three newly designed oligonucleotide primers specific to the variable lipoprotein haemagglutinin (vlhA) gene and differentiate M. synoviae field strains based on a nucleotide deletion or the insertion of the proline-rich repeat (PRR) region of the vlhA gene. The developed semi-nested polymerase chain reaction (PCR) assay revealed positive results in 12 out of 100 clinical samples collected from chickens showing lameness and joint swelling. Six positive samples were selected randomly for sequencing, and sequence analysis revealed 96.3-100% nucleotide identities compared to the reference sequences. Phylogenetic analysis showed that sequences of the strains in this study were closely related to WVU1853 (Spain), CK.MS.UDL.PK.2014.2 (Pakistan), and F10-2AS (USA) strains, but they were distinct from the M. synoviae-H vaccine strain sequence. M. synoviae obtained from these samples were identified as types A and C with a length of 38 and 32 amino acids, respectively. These results indicated that the specific and sensitive semi-nested PCR could be a useful diagnostic tool for the direct identification of clinical samples, and the sequence analysis of the partial vlhA gene can be useful for typing M. Synoviae.

Identification and Phylogeny of the Human Endogenous Retrovirus HERV-W LTR Family in Schizophrenia

  • Huh, Jae-Won;Yi, Joo-Mi;Kim, Heui-Soo
    • Journal of Life Science
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    • v.11 no.2
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    • pp.83-86
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    • 2001
  • The long terminal repeat (LTR) elements of human endogenous retrovirus (HERV) have been found to be coexpressed with genes located nearby. It has been suggested that the LTR elements have contributed to the genetic variation of human genome connected to various diseases. Recently, HERV-W family was identified in the cerebrospinal fluids and brains of individuals with schizophrenia. Using genomic DNAs derived from schizophrenia, we performed PCR amplification and identified six HERV-W LTR elements. Those LTR elements showed a high degree of sequence similarity (87.7-99.5%) with HERV-W LTR (AF072500). Sequence analysis of the HERV-W LTR elements revealed that clone W-sch1 showed identical sequence with the AC003014 (PAC clone RP1-290B4) derived from human Xq23. Clone W-sch2 was closely related to the AC0072442 derived from human Y chromosome by phylogenetic analysis. Our data suggest that new HERV-W LTR elements in schizophrenia may be very useful for further studies to understand neuropsychiatric diseases.

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