Production of polyclonal antibodies against recombinant potato virus X coat protein
Čeřovská, Noemi; Plchová, Helena; Moravec, Tomáš; Filigarová, Marie; Dědič, P.
2008 - English
The gene encoding the coat protein (CP) of Potato virus X was cloned into expression vector pMPM-4A Omega. PVX CP was expressed in Escherichia coli and was used for raising rabbit polyclonal antibodies. For immunization we purified CP fractions from bacterial lysate either by simple fractionation or by excision from SDS gel. The obtained sera and antibodies were tested for the detection of PVX in laboratory hosts Nicotiana tabacum, cv. Petit Havana SR 1 and N.benthamiana and in various cultivars of natural host Solanum tuberosum by ELISA as well as by Western blots. The antibodies were specific for PVX and are usable for the detection of the broad isolates spectrum of the Potato virus X by ELISA and Western blot. Gen kódující obalový protein (CP) X viru bramboru (PVX) byl klónován v expresním vektoru pMPM-4AΩ . PVX CP byl exprimován v Escherichia coli a byl použit pro produkci králičích polyklonálních protilátek.Získaná antiséra a protilátky byly testovány na přítomnost PVX v hostitelských rostlinách Nicotiana tabacum cv. Petit Havana SR 1 a N. benthamiana a v různých kultivarech přírodního hostitele Solanum tuberosum Protilátky byly specifické pro PVX a použitelné pro detekci širokého spektra izolátů PVX při použití metody IPTA ELISA a Western blotu.
Keywords:
polyclonal antibodies; recombinant coat protein; Y potato virus
Available at various institutes of the ASCR
Production of polyclonal antibodies against recombinant potato virus X coat protein
The gene encoding the coat protein (CP) of Potato virus X was cloned into expression vector pMPM-4A Omega. PVX CP was expressed in Escherichia coli and was used for raising rabbit polyclonal ...
Chromosome-based approaches for physical mapping in wheat
Šafář, Jan; Bartoš, Jan; Šimková, Hana; Doleželová, Marie; Janda, Jaroslav; Kovářová, Pavlína; Suchánková, Pavla; Doležel, Jaroslav
2005 - English
Keywords:
Physical map; chromosome; wheat
Available at various institutes of the ASCR
Chromosome-based approaches for physical mapping in wheat
Different interactions between cytokinins and abscisic acid
Pospíšilová, Jana; Baťková, Petra; Vágner, Martin; Malbeck, Jiří; Trávníčková, Alena
2005 - English
Keywords:
abscisic acid; N6-benzyladenine
Available at various institutes of the ASCR
Different interactions between cytokinins and abscisic acid
The impact of constitutive overexpression of trans-zeatin O-glucosyltransferase gene in tobacco on photosynthetic pigments and gas exchange during water stress
Haisel, Daniel; Pospíšilová, Jana; Nováková, Marie; Vaňková, Radomíra
2005 - English
Keywords:
trans-zeatin; water stress
Available at various institutes of the ASCR
The impact of constitutive overexpression of trans-zeatin O-glucosyltransferase gene in tobacco on photosynthetic pigments and gas exchange during water stress
Změny ve fotosyntetických charakteristikách listů tabáku ve dvou stádiích infekce TMV
Procházková, Dagmar; Wilhelmová, Naděžda; Šindelářová, Milada; Šindelář, Luděk
2005 - English
Available at various institutes of the ASCR
Změny ve fotosyntetických charakteristikách listů tabáku ve dvou stádiích infekce TMV
Evaluation of the possible role of cytokinin O-glucosylation in response of tobacco plants to drought stress
Nováková, Marie; Dobrev, Petre; Motyka, Václav; Gaudinová, Alena; Malbeck, Jiří; Pospíšilová, Jana; Martin, R. C.; Mok, D. W. S.; Mok, M. C.; Vaňková, Radomíra
2005 - English
Keywords:
cytokinins; o-glucosyltranferase
Available at various institutes of the ASCR
Evaluation of the possible role of cytokinin O-glucosylation in response of tobacco plants to drought stress
Trichloroacetic acid in Norway spruce: Aspects of plant uptake
Forczek, Sándor; Albrechtová, Jolana; Matucha, Miroslav; Uhlířová, H.
2004 - English
Keywords:
cytokinins; therapy
Available at various institutes of the ASCR
Trichloroacetic acid in Norway spruce: Aspects of plant uptake
Dissecting large genomes of Triticeae by chromosome sorting
Doležel, Jaroslav; Kubaláková, Marie; Suchánková, Pavla; Šafář, Jan; Janda, Jaroslav; Kovářová, Pavlína; Bartoš, Jan; Čihalíková, Jarmila; Šimková, Hana
2004 - English
Nuclear genomes of some Triticeae species are characterized by large size and prevalence of repetitive DNA sequences. These features hamper their physical mapping and gene cloning. Purification of individual chromosomes by flow cytometry can simplify these tasks by providing small and defined genome fractions. This lecture reviews the development of the methodology and its potential for genome mapping in barley, rye and wheat Due to small differences in relative DNA content, only one chromosome type can be discriminated and sorted in each of the three species. Cytogenetic stocks facilitate separation of other parts of the genomes as individual chromosomes, translocation chromosomes and chromosome arms. Chromosome analysis by flow cytometry permits quantitative detection of structural and numerical chromosome changes. Chromosomes sorted onto microscopic slides have been used for discovery of rare structural changes and for high-resolution cytogenetic mapping using FISH. The use of sorted chromosomes for HAPPY mapping, targeted isolation of low copy "genie" sequences, and high-throughput physical mapping of ESTs on microarrays are attractive options. As millions of chromosomes with intact DNA may be sorted, construction of BAC libraries is possible. Subgenomic, chromosome-specific and chromosome arm-specific BAC libraries have already been produced in wheat and represent unique resources for genomics of cereals. Čeleď Triticeae je charakteristická velkým obsahem DNA repetitivní povahy. Tento fakt komplikuje jejich fyzické mapování a klonování genů. Izolace jednotlivých chromozomů průtokovou cytometrií umožňuje zjednodušit a zrychlit studium těchto genomů. Tříděných chromozomů může být využito pro HAPPY mapování, FISH techniky, mapování EST sekvencí, konstrukci chromozomově specifických BAC knihoven a řadě dalších.
Keywords:
Triticeae sp.; flow cytogenetics; FISH
Available at various institutes of the ASCR
Dissecting large genomes of Triticeae by chromosome sorting
Nuclear genomes of some Triticeae species are characterized by large size and prevalence of repetitive DNA sequences. These features hamper their physical mapping and gene cloning. Purification of ...
Advances in flow cytogenetics of barley
Suchánková, Pavla; Kubaláková, Marie; Kovářová, Pavlína; Bartoš, Jan; Čihalíková, Jarmila; Endo, T. R.; Doležel, Jaroslav
2004 - English
Flow cytogenetics is an attractive tool that facilitates genome mapping. Flow cytometric analysis permits classification of isolated chromosomes according to their DNA content As thousands of chromosomes are analysed in each sample, quantitative detection of structural and numerical chromosome changes is possible. Any chromosome that can be discriminated can also be sorted in large quantities and high purities for subsequent molecular analysis. Unfortunately, only the smallest chromosome 5(1H) can be discriminated and sorted in barley. This is due to similarity in relative DNA content among the chromosomes 1 - 4,6 and 7 (2H - 7H). Cytogenetic stocks facilitate sorting of those translocation chromosomes that differ in DNA content Even if the sorted translocation chromosomes were found useful for mapping DNA sequences to subchromosomal regions, their use remained limited. Here we describe a novel approach that facilitates sorting of any of the 14 barley chromosome arms. The strategy is based on using wheat-barley addition lines carrying barley chromosome arms (telocentric chromosomes). Furthermore, we will be able to sort subarm chromosomal segments that are generated in the wheat-barley chromosome addition lines by the gametocidal system. These advances make flow cytogenetics an attractive tool that can simplify barley genome mapping. A possibility to purify large quantities of individual chromosome arms and their segments opens avenues for targeted isolation of low-copy "genie" sequences, preparation of specific probes for screening EST arrays, HAPPY mapping, and construction of chromosome arm-specific BAG libraries. Analýza pomocí průtokového cytometru umožňuje klasifikaci izolovaných chromozómů podle obsahu DNA. Každý chromozóm, který může být pomocí průtokového cytometru identifikován, může být zároveň velkou rychlostí tříděn z populace všech chromozómů. U ječmene však může být tříděn pouze nejmenší chromozóm 1H. Nová strategie je založena na použití adičních linií pšenice-ječmen nesoucích v genomu pšenice chromozómová ramena ječmene ve formě telocentrických chromozómů. Tato strategie umožňuje třídění jakéhokoliv ze 14 chromozómových ramen ječmene.
Keywords:
Flow cytogenetics; barley; flow karytyping
Available at various institutes of the ASCR
Advances in flow cytogenetics of barley
Flow cytogenetics is an attractive tool that facilitates genome mapping. Flow cytometric analysis permits classification of isolated chromosomes according to their DNA content As thousands of ...
Quantitative detection of mRNA in tobacco
Štorchová, Helena; Libus, Jiří; Vaňková, Radomíra
2004 - English
Keywords:
Quantitative RT PCR; optimization; tobacco
Available at various institutes of the ASCR
Quantitative detection of mRNA in tobacco
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