A Simple Protocol for the Expression and Purification of NC2
Transcriptional regulation depends on the appropriate set of positive and negative regulating signals in order to provide the correct gene expression. Studies in eukaryotic gene expression over the last few years have shown that NC2 acts as a general repressor of transcription. Functional in vitro studies require large amounts of highly purified recombinant protein. The case of rNC2a (Drap1) which up to date has been difficult to express in bacteria is common among many eukaryotic proteins. Moreover, under current protocols both subunits have to be denature in urea after combination and allow a renaturation process which may render many inactive NC2 complexes. Here we provide a simple protocol that includes a new expression vector and the protocol for over expressing and purifying both subunits in a native state.
| Main Authors: | , , |
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| Format: | info:eu-repo/semantics/article biblioteca |
| Language: | eng |
| Subjects: | info:eu-repo/classification/Autores/DRL, info:eu-repo/classification/Autores/DRAPL, info:eu-repo/classification/Autores/NC2, info:eu-repo/classification/Autores/REPRESSOR, info:eu-repo/classification/Autores/TRANSCRIPTION, info:eu-repo/classification/Autores/GENE REGULATION, info:eu-repo/classification/cti/2, |
| Online Access: | http://cicy.repositorioinstitucional.mx/jspui/handle/1003/798 |
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| Summary: | Transcriptional regulation depends on the appropriate set of positive and negative regulating signals in order to provide the correct gene expression. Studies in eukaryotic gene expression over the last few years have shown that NC2 acts as a general repressor of transcription. Functional in vitro studies require large amounts of highly purified recombinant protein. The case of rNC2a (Drap1) which up to date has been difficult to express in bacteria is common among many eukaryotic proteins. Moreover, under current protocols both subunits have to be denature in urea after combination and allow a renaturation process which may render many inactive NC2 complexes. Here we provide a simple protocol that includes a new expression vector and the protocol for over expressing and purifying both subunits in a native state. |
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