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Isolation and analysis of mRNAs from specific cell types of plants by ribosome immunopurification

Title data

Mustroph, Angelika ; Zanetti, Maria Eugenia ; Girke, Thomas ; Bailey-Serres, Julia:
Isolation and analysis of mRNAs from specific cell types of plants by ribosome immunopurification.
In: Smet, Ive de (ed.): Plant organogenesis : methods and protocols. - New York : Humana Press , 2013 . - pp. 277-302 . - (Methods in Molecular Biology ; 959 )
ISBN 978-1-62703-220-9
DOI: https://doi.org/10.1007/978-1-62703-221-6_19

Official URL: Volltext

Abstract in another language

Multiple ribosomes assemble onto an individual mRNA to form a polyribosome (polysome) complex. The epitope tagging of specific ribosomal proteins can enable the immunopurification of polysomes from crude cell extracts derived from cryopreserved tissue samples. Through expression of the epitope-tagged ribosomal protein in cell-type and regional specific domains of Arabidopsis thaliana and other organisms it is feasible to quantitatively assess the mRNAs that are associated with ribosomes with cell-specific resolution. Here we present detailed methods for development of transgenics that express a FLAG-tagged version of ribosomal protein L18 (RPL18) under the direction of individual promoters with specific domains of expression, the immunopurification of ribosomes, and bioinformatic analyses of the resultant datasets obtained by microarray profiling. This methodology provides researchers with the opportunity to assess rapid changes at the organ, tissue, regional or cell-type specific level of mRNAs that are associated with ribosomes and therefore engaged in translation.

Further data

Item Type: Article in a book
Refereed: Yes
Additional notes: PubMed-ID: 23299683
BAYCEER116716
Institutions of the University: Faculties > Faculty of Biology, Chemistry and Earth Sciences
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Biology
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Biology > Professorship Plant Genetics
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Biology > Professorship Plant Genetics > Professorship Plant Genetics- Univ.Prof. Dr. Angelika Mustroph
Profile Fields
Profile Fields > Advanced Fields
Profile Fields > Advanced Fields > Molecular Biosciences
Research Institutions
Research Institutions > Research Centres
Research Institutions > Research Centres > Bayreuth Center for Molecular Biosciences - BZMB
Research Institutions > Research Centres > Bayreuth Center of Ecology and Environmental Research- BayCEER
Faculties
Result of work at the UBT: No
DDC Subjects: 500 Science > 570 Life sciences, biology
500 Science > 580 Plants (Botany)
Date Deposited: 01 Apr 2015 06:49
Last Modified: 03 Jul 2015 09:09
URI: https://eref.uni-bayreuth.de/id/eprint/9611