• Title/Summary/Keyword: RNA mapping

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Some Properties of an Isolate of Peanut stunt virus Isolated from White Clover (Trifolium repens L.) (토끼풀에서 분리한 Peanut stunt virus의 성질)

  • Jung, Goo-Ho;Jeon, Yong-Woon;Choi, Jang-Kyung;Hong, Jin-Sung;Ryu, Ki-Hyun;Lee, Sang-Yong
    • Research in Plant Disease
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    • v.14 no.1
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    • pp.71-75
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    • 2008
  • An isolate of Peanut stunt virus (PSV), named as Tr-PSV, was isolated from white clover (Trifolium repens L) showing mosaic symptom. Tr-PSV systemically infected all plants tested in the Nicotiana spp. and induced local lesions on inoculated leaves of Chenopodium amaranticolor. However, Tr-PSV induced typical mosaic symptoms as ER-PSV on Vigna unguiculata 5 to 6 days after inoculation, while Fny-CMV used as a control virus of Cucumovirus produced local lesions on inoculated leaves. In dsRNA analysis, Tr-PSV consisted of four dsRNAs, but satellite RNA was not detected. The cDNA of coat protein gene of Tr-PSV was amplified by RT-PCR using a Cucumovirus-specific single pair primers that designed to amplify a DNA fragment of approximately 950 bp. By restriction mapping analysis using RFLP of the RT-PCR products and by serological properties of gel diffusion test, Tr-PSV belongs to a typical member of PSV subgroup I. This is the first report on the occurrence of PSV in white clover in Korea.

Characterization and Sequence Analysis of a Lily Isolate of Cucumber mosaic virus from Lithium tsingtauense

  • Ryu, Ki-Hyun;Park, Hye-Won;Park, Jang-Kyung
    • The Plant Pathology Journal
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    • v.18 no.2
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    • pp.85-92
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    • 2002
  • A new isolate of Cucumber mosaic virus (CMV), identified as Li-CMV was isolated from a diseased Korean native lily (Lithium tsingtauense Gilg). Biological and serological properties of Li-CMV were characterized, and reverse transcription-polymerase chain reaction (RT-PCR) analysis, restriction enzyme profiling of RT-PCR products, and nucleotide sequence analysis of RNA3 of the virus were performed in this study. Remarkable differences in symptoms between Li-CMV and ordinary CMV strains were found in tobacco plants and Datura stramonium. Li-CMV-infected tobacco plants (cv. Xanthi-nc and cv. Samsun) induced chlorotic ringspots on uninoculated upper leaves, and the symptom expression was delayed or faint whereas, ordinary CMV strains induced green mosaic symptoms on the plant. Systemic infections were observed on Nicotiana benthamiana with severe mosaic symptom. Restriction mapping analysis of RT-PCR products using MspI showed that Li-CMV belonged to CMV subgroup I. A full-length CDNA copy of RNA3 for the virus was amplified by RT-PCR, cloned, and its complete nucleotide sequence was determined. The RNA3 of Li-CMV was 2, 232 nucleotides long, and consisted of two open reading frames of 843 and 657 bases encoding 3a protein (movement protein) and coat protein, respectively. Results of this study indicate that Li-CMV is a novel strain and belongs to subgroup I of CMV in the genus Cucumovirus.

Genome Mapping of an Extreme Thermophile, Thermus caldophilus GK24

  • Park, Jong Hoon;Park, Byung Chul;Koch, Suk Hoon;Kim, Joong Soo;Koh, Jeong Heon;Yang, Moon Hee;Kim, Yong Sung;Kim, Cheorl Ho;Kim, Myoung Hee;Kwon, Suk Tae;Lee, Dae-Sil
    • Genomics & Informatics
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    • v.1 no.1
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    • pp.50-54
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    • 2003
  • Genome of an extreme thermophile, Thermus caldophilus GK24 has been analyzed to construct the genomic map. The genomic DNAs encapsulated in agarose gel were digested with SspI, EcoRI, SpeI, and HpaI restriction endonucleases, and then the resulting genomic DNA fragments were analyzed by pulsed-field gel electrophoresis. Its restriction map has been constructed by analyzing sizes of the restriction fragments obtained from both complete and partial digestions. The circular form of its genome was composed of about 1.98 Mbp and a megaplasmid. The genomic loci for the genes of xylose isomerase, thioredoxin, tRNA-16S rRNA, 23S rRNA, L5 ribosomal protein, ADP-glucose pyrophosphorylase, DNA-ligase, and Tca DNA polymerase were determined by both Southern hybridization and PCR.

Investigation of Splicing Quantitative Trait Loci in Arabidopsis thaliana

  • Yoo, Wonseok;Kyung, Sungkyu;Han, Seonggyun;Kim, Sangsoo
    • Genomics & Informatics
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    • v.14 no.4
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    • pp.211-215
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    • 2016
  • The alteration of alternative splicing patterns has an effect on the quantification of functional proteins, leading to phenotype variation. The splicing quantitative trait locus (sQTL) is one of the main genetic elements affecting splicing patterns. Here, we report the results of genome-wide sQTLs across 141 strains of Arabidopsis thaliana with publicly available next generation sequencing datasets. As a result, we found 1,694 candidate sQTLs in Arabidopsis thaliana at a false discovery rate of 0.01. Furthermore, among the candidate sQTLs, we found 25 sQTLs that overlapped with the list of previously examined trait-associated single nucleotide polymorphisms (SNPs). In summary, this sQTL analysis provides new insight into genetic elements affecting alternative splicing patterns in Arabidopsis thaliana and the mechanism of previously reported trait-associated SNPs.

A new serotype confirmed by partial physical mapping of cDNA clones from the infectious pancreatic necrosis virus (IPNV) isolated in Korea (한국에서 분리된 전염성 췌장괴저 바이러스의 새로운 혈청형에 대한 유전자 분석)

  • 이정진;박정우;정가진
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.231-236
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    • 1989
  • The larger segment of double stranded RNA genome from a new serotype of Infectious Pancreatic Necrosis Virus (IPNV), DRT, has been partially cloned at Sma I site in pUC19 and compared with the restriction maps of VR-299 and Sp. Restrction sites found in DRT was distinct and hence a new serotype. The cDNA clones of DRT were about 800, 850, and 1, 400 bp long each and do not share any common restiction site. It is not clear yet if there exist any overlapping sequences among them. This partial cloning, however, was sufficient for the comparison of restriction maps with the other serotypes.

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Protein Interaction Mapping of Translational Regulators Affecting Expression of the Critical Stem Cell Factor Nos

  • Malik, Sumira;Jang, Wijeong;Kim, Changsoo
    • Development and Reproduction
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    • v.21 no.4
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    • pp.449-456
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    • 2017
  • The germline stem cells of the Drosophila ovary continuously produce eggs throughout the life-span. Intricate regulation of stemness and differentiation is critical to this continuous production. The translational regulator Nos is an intrinsic factor that is required for maintenance of stemness in germline stem cells. Nos expression is reduced in differentiating cells at the post-transcriptional level by diverse translational regulators. However, molecular mechanisms underlying Nos repression are not completely understood. Through three distinct protein-protein interaction experiments, we identified specific molecular interactions between translational regulators involved in Nos repression. Our findings suggest a model in which protein complexes assemble on the 3' untranslated region of Nos mRNA in order to regulate Nos expression at the post-transcriptional level.

Characterization of an Isolate of Cucumber mosaic virus from Raphanus sativus L. (열무에서 분리한 오이모자이크바이러스 분리주의 특성)

  • Rhee, Sun-Ju;Hong, Jin-Sung;Choi, Jang-Kyung;Kim, Eun-Ji;Lee, Gung-Pyo
    • Research in Plant Disease
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    • v.17 no.2
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    • pp.211-215
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    • 2011
  • Cucumber mosaic virus (CMV)-like isolate was collected from Raphanus sativus (cv. Choon-hyang), which showed mosaic symptoms. The isolate was confirmed to a strain of CMV by host responses in Vigna unguiculata, Chenopodium amaranticolor and Gomphrena globosa, by viral genome composition with RT-PCR and PCR-RFLP, and by serological analysis. Symptom developed by the strain of CMV was severe in Nicotiana benthamiana, N. glutinosa, N. tabacum (cv. Samsun, cv. Xanthi), Cucumis melo (cv. Early hanover), Cucumis sativus (cv. White wonder), Capsicum annuum (cv. Chung-yang and cv. Geum-top), but mild symptom was developed in Raphanus sativus (cv. Choon-hyang), Brassica rapa ssp. pekinensis (cv. Bul-Am No. 3), and B. juncea (cv. Daenong Jukgot). Newly isolated strain of CMV could infect diverse crops including Solanaceae, Cucurbitaceae and Brassicaceae. We designated the new strain of CMV as Gn-CMV based on the novel infectivity of Brassicaceae. In double-stranded (ds) RNA analysis, Gn-CMV consisted of 3.3, 3.0, and 2.2 kb genomes likewise other strains of CMV. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) showed 28 kDa of the CMV coat protein. By restriction enzyme mapping using Cac8I, ClaI and MspI of RT-PCR products indicated that Gn-CMV belongs to CMV subgroup I.

Screening for candidate genes related with histological microstructure, meat quality and carcass characteristic in pig based on RNA-seq data

  • Ropka-Molik, Katarzyna;Bereta, Anna;Zukowski, Kacper;Tyra, Miroslaw;Piorkowska, Katarzyna;Zak, Grzegorz;Oczkowicz, Maria
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.10
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    • pp.1565-1574
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    • 2018
  • Objective: The aim of the present study was to identify genetic variants based on RNA-seq data, obtained via transcriptome sequencing of muscle tissue of pigs differing in muscle histological structure, and to verify the variants' effect on histological microstructure and production traits in a larger pig population. Methods: RNA-seq data was used to identify the panel of single nucleotide polymorphisms (SNPs) significantly related with percentage and diameter of each fiber type (I, IIA, IIB). Detected polymorphisms were mapped to quantitative trait loci (QTLs) regions. Next, the association study was performed on 944 animals representing five breeds (Landrace, Large White, Pietrain, Duroc, and native Puławska breed) in order to evaluate the relationship of selected SNPs and histological characteristics, meat quality and carcasses traits. Results: Mapping of detected genetic variants to QTL regions showed that chromosome 14 was the most overrepresented with the identification of four QTLs related to percentage of fiber types I and IIA. The association study performed on a 293 longissimus muscle samples confirmed a significant positive effect of transforming acidic coiled-coil-containing protein 2 (TACC2) polymorphisms on fiber diameter, while SNP within forkhead box O1 (FOXO1) locus was associated with decrease of diameter of fiber types IIA and IIB. Moreover, subsequent general linear model analysis showed significant relationship of FOXO1, delta 4-desaturase, sphingolipid 1 (DEGS1), and troponin T2 (TNNT2) genes with loin 'eye' area, FOXO1 with loin weight, as well as FOXO1 and TACC2 with lean meat percentage. Furthermore, the intramuscular fat content was positively associated (p<0.01) with occurrence of polymorphisms within DEGS1, TNNT2 genes and negatively with occurrence of TACC2 polymorphism. Conclusion: This study's results indicate that the SNP calling analysis based on RNA-seq data can be used to search candidate genes and establish the genetic basis of phenotypic traits. The presented results can be used for future studies evaluating the use of selected SNPs as genetic markers related to muscle histological profile and production traits in pig breeding.

Karyotype Analysis and rDNA Physical Mapping in Rye (Secale cereale L.) (호밀(Secale cereale L.)의 핵형분석과 rDNA의 Physical Mapping)

  • Lee, Joon Soo;Seo, Bong Bo;Kim, Min
    • Korean Journal of Breeding Science
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    • v.42 no.2
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    • pp.163-168
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    • 2010
  • This study was carried out to determine the chromosomal localization of the 5S and 18S-26S ribosomal DNA(rDNA) genes by means of fluorescence in situ hybridization(FISH) techniques, and the constitutive heterochromatin detected by means of Gimsa C-banding technique in rye(Secale cereale L.). The somatic chromosomes number was 2n=14. The karyotype consists of four pairs of metacentrics(chromosomes 1, 2, 3, and 7) and three pairs of submetacentrics(chromosomes 4, 5, and 6). Secondary constrictions appeared in the short arm of chromosome 1. The 5S rDNA genes have been located on two pairs of chromosomes 1 and 5, and 18S-26S rDNAs genes have been located on one pair of chromosome 1. 5S rDNA genes were detected on the distal region of the secondary constrictions in nucleolus organizer regions(NOR) in chromosome 1, and other detected on the intercalary region in the short arm of chromosome 5.

Identification and Characterization of Three Isolates of Cucumber mosaic virus Isolated from Weed Hosts (잡초에서 분리한 3종 Cucumber mosaic virus의 동정과 특성)

  • Lee, Hyeok-Geun;Kim, Sung-Ryul;Jeon, Yong-Woon;Kwon, Soon-Bae;Ryu, Ki-Hyun;Choi, Jang-Kyung
    • Research in Plant Disease
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    • v.14 no.1
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    • pp.15-20
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    • 2008
  • Three isolates of Cucumber mosaic virus (CMV) were isolated from weed hosts showing typical mosaic symptoms, and some properties of the viruses were investigated. CMV isolates, designated as Is-CMV, Jd-CMV and Pla-CMV from Isodon inflexus, Jeffersonia dubia and Phryma leptostachya var. asiatica, respectively, were identified and characterized by biological reaction in several host plants, serological property, dsRNA analysis, reverse transcription-polymerase chain reaction (RT-PCR), restriction fragment-length polymorphism (RFLP). All isolates systemically infected in Nicotiana benthamiana, Cucurbita pepo cv. Black beauty and Cucumis sativus, and did not reveal any differences in these host plants between the isolates. However, remarkable difference in the symptoms was found between the CMVs in Capsicum annuum. Is-CMV induced an asymptomatic symptoms, while Jd-CMV and Pla-CMV produced severe mosaic symptoms in C. annuum plants. In dsRNA analysis, all isolates revealed four major bands with estimated molecular size of 3.4, 3.2, 2.1 and 1.0 kbp. The cDNAs of coat protein gene of the isolates were amplified by RT-PCR using a genus-specific single pair primers that designed to amplify a DNA fragment of approximately ranging from 938 to 966 bp. By restriction mapping analysis using RFLP of the RT-PCR products as well as by serological properties of gel diffusion test, the CMV isolates belong to a typical members of CMV subgroup IA. This is the first report on the occurrence of CMV in the three weed hosts.