Chapter title |
Isolation of Polysomal RNA for Analyzing Stress-Responsive Genes Regulated at the Translational Level in Plants
|
---|---|
Chapter number | 9 |
Book title |
Plant Stress Tolerance
|
Published in |
Methods in molecular biology, July 2017
|
DOI | 10.1007/978-1-4939-7136-7_9 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7134-3, 978-1-4939-7136-7
|
Authors |
Yong-Fang Li, Ramamurthy Mahalingam, Ramanjulu Sunkar, Li, Yong-Fang, Mahalingam, Ramamurthy, Sunkar, Ramanjulu |
Abstract |
Alteration of gene expression is an essential mechanism, which allows plants to respond and adapt to adverse environmental conditions. Transcriptome and proteome analyses in plants exposed to abiotic stresses revealed that protein levels are not correlated with the changes in corresponding mRNAs, indicating regulation at translational level is another major regulator for gene expression. Analysis of translatome, which refers to all mRNAs associated with ribosomes, thus has the potential to bridge the gap between transcriptome and proteome. Polysomal RNA profiling and recently developed ribosome profiling (Ribo-seq) are two main methods for translatome analysis at global level. Here, we describe the classical procedure for polysomal RNA isolation by sucrose gradient ultracentrifugation followed by highthroughput RNA-seq to identify genes regulated at translational level. Polysomal RNA can be further used for a variety of downstream applications including Northern blot analysis, qRT-PCR, RNase protection assay, and microarray-based gene expression profiling. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 2 | 100% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 2 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 8 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 2 | 25% |
Student > Ph. D. Student | 1 | 13% |
Student > Doctoral Student | 1 | 13% |
Unknown | 4 | 50% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 2 | 25% |
Agricultural and Biological Sciences | 2 | 25% |
Unknown | 4 | 50% |