Isolation of RNP granules
Publikation: Bidrag til bog/antologi/rapport › Bidrag til bog/antologi › Forskning › fagfællebedømt
The post-transcriptional operon provides a means of synexpression of mRNAs encoding interrelated proteins. The coordination of gene expression may be achieved by a trans-acting RNA-binding protein attaching to similar cis-elements in different, yet functionally clustered, mRNAs. The RNP granule can be regarded as a supramolecular assembly of RNA and protein, probably representing several overlapping post-transcriptional operons. The present protocol describes how RNP granules may be isolated by the transgenic expression of a 3X FLAG version of an RNA-binding protein under tetracycline control via the tetracycline receptor/operator complex. In this way, inclusion of an appropriate tetracycline concentration ensures expression of the tagged version at the endogenous level, and the 3X FLAG tag is a convenient "handle" for the subsequent immunoprecipitation by immobilized anti-FLAG antibody.
Originalsprog | Engelsk |
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Titel | RNA : methods and protocols |
Redaktører | Henrik Nielsen |
Antal sider | 9 |
Vol/bind | 703 |
Forlag | Springer |
Publikationsdato | 2011 |
Sider | 265-273 |
Kapitel | 18 |
ISBN (Trykt) | 978-1-58829-913-0 |
ISBN (Elektronisk) | 978-1-59745-248-9 |
DOI | |
Status | Udgivet - 2011 |
Navn | Methods in Molecular Biology |
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ISSN | 1064-3745 |
ID: 40966820