Establishment of a Cre/loxP recombination system for N-terminal epitope tagging of genes in <it>Tetrahymena</it>
<p>Abstract</p> <p>Background</p> <p>Epitope tagging is a powerful strategy to study the function of proteins. Although tools for C-terminal protein tagging in the ciliated protozoan <it>Tetrahymena thermophila </it>have been developed, N-terminal protein ta...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2010-07-01
|
| Series: | BMC Microbiology |
| Online Access: | http://www.biomedcentral.com/1471-2180/10/191 |
| Summary: | <p>Abstract</p> <p>Background</p> <p>Epitope tagging is a powerful strategy to study the function of proteins. Although tools for C-terminal protein tagging in the ciliated protozoan <it>Tetrahymena thermophila </it>have been developed, N-terminal protein tagging in this organism is still technically demanding.</p> <p>Results</p> <p>In this study, we have established a Cre/loxP recombination system in <it>Tetrahymena </it>and have applied this system for the N-terminal epitope tagging of <it>Tetrahymena </it>genes. Cre can be expressed in <it>Tetrahymena </it>and localizes to the macronucleus where it induces precise recombination at two loxP sequences in direct orientation in the <it>Tetrahymena </it>macronuclear chromosome. This Cre/loxP recombination can be used to remove a loxP-flanked drug-resistance marker from an N-terminal tagging construct after it is integrated into the macronucleus.</p> <p>Conclusions</p> <p>The system established in this study allows us to express an N-terminal epitope tagged gene from its own endogenous promoter in <it>Tetrahymena</it>.</p> |
|---|---|
| ISSN: | 1471-2180 |