Localisation of abundant and organ-specific genes expressed in Rosa hybrida leaves and flower buds by direct in situ RT-PCR

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Jedrzejuk, A.; Mibus, H.; Serek, M.: Localisation of abundant and organ-specific genes expressed in Rosa hybrida leaves and flower buds by direct in situ RT-PCR. In: The Scientific World Journal 2012 (2012), 609597. DOI: https://doi.org/10.1100/2012/609597

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Sum total of downloads: 166




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Abstract: 
In situ PCR is a technique that allows specific nucleic acid sequences to be detected in individual cells and tissues. In situ PCR and IS-RT-PCR are elegant techniques that can increase both sensitivity and throughput, but they are, at best, only semiquantitative; therefore, it is desirable first to ascertain the expression pattern by conventional means to establish the suitable conditions for each probe. In plants, in situ RT-PCR is widely used in the expression localisation of specific genes, including MADS-box and other function-specific genes or housekeeping genes in floral buds and other organs. This method is especially useful in small organs or during early developmental stages when the separation of particular parts is impossible. In this paper, we compared three different labelling and immunodetection methods by using in situ RT-PCR in Rosa hybrida flower buds and leaves. As target genes, we used the abundant -actin and RhFUL gene, which is expressed only in the leaves and petals/sepals of flower buds. We used digoxygenin-11-dUTP, biotin-11-dUTP, and fluorescein-12-dUTP-labelled nucleotides and antidig-AP/ streptavidin- fluorescein-labelled antibodies. All of the used methods gave strong, specific signal and all of them may be used in localization of gene expression on tissue level in rose organs.
License of this version: CC BY 3.0 Unported
Document Type: Article
Publishing status: publishedVersion
Issue Date: 2012
Appears in Collections:Naturwissenschaftliche Fakultät

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pos. country downloads
total perc.
1 image of flag of Germany Germany 111 66.87%
2 image of flag of United States United States 22 13.25%
3 image of flag of China China 20 12.05%
4 image of flag of No geo information available No geo information available 2 1.20%
5 image of flag of Russian Federation Russian Federation 1 0.60%
6 image of flag of Netherlands Netherlands 1 0.60%
7 image of flag of Italy Italy 1 0.60%
8 image of flag of India India 1 0.60%
9 image of flag of Hungary Hungary 1 0.60%
10 image of flag of Canada Canada 1 0.60%
    other countries 5 3.01%

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