{"id":1813,"date":"2021-06-05T16:48:22","date_gmt":"2021-06-05T16:48:22","guid":{"rendered":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/?page_id=1813"},"modified":"2022-01-25T16:58:35","modified_gmt":"2022-01-25T16:58:35","slug":"mammalian-expression-vectors","status":"publish","type":"page","link":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/services\/strategy-and-construct-design\/mammalian-expression-vectors\/","title":{"rendered":"Mammalian expression vectors"},"content":{"rendered":"\n
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Overview of our mammalian expression vectors<\/a><\/p>\n\n\n\n

For transient expression of single proteins in mammalian cells, we mainly use the pXLG vector, which was a kind gift from David Hacker from the EPFL (Switzerland).<\/p>\n\n\n\n

We also have a collection of mammalian expression vectors specifically adapted for ligation-independent cloning. These vectors are called pCoofy<\/a><\/strong> vectors and were developed by Sabine Suppmann at the Max Planck Institute for Biochemistry in Martinsried, Munich.<\/p>\n\n\n\n

The detailed cloning protocols and descriptions of these vectors can be found in the following reference:<\/p>\n\n\nScholz J., Besir H., Strasser S. and Suppmann S. (2013) A new method to customize protein expression vectors for fast, efficient and background free parallel cloning. BMC Biotechnology<\/em>. 13<\/strong>:12<\/blockquote>\n\n\n\n

Internal EMBL users can obtain these vectors at the Protein Expression and Purification Core Facility. External users can request them via Addgene<\/a>.<\/p>\n\n\n\n

Mammalian pCOOFY expression vectors<\/h3>\n\n\n\n
Name<\/th>N-terminal tag<\/th>C-terminal tag<\/th>Antibiotic resistance<\/th>Backbone<\/th><\/tr><\/thead>
pCoofy47<\/td>VEGFss-3C<\/td>* His10<\/sub>
* TwinStrepII
* S-tag
* CBP<\/td>		Ampicillin<\/td>pTT5<\/td><\/tr>
pCoofy50<\/td>HA-His8<\/sub>-Flag-3C<\/td>His10<\/sub><\/td>Ampicillin<\/td>pEF-HA<\/td><\/tr>
pCoofy55<\/td>His10<\/sub>-SumoStar-3C<\/td>* His10<\/sub>
* TwinStrepII
* S-tag<\/td>		Ampicillin<\/td>pTT5<\/td><\/tr>
pCoofy56<\/td>TwinStrepII-3C<\/td>His10<\/sub><\/td>Ampicillin<\/td>pEF-HA<\/td><\/tr>
pCoofy57<\/td>VEGFss-His8<\/sub>-3C<\/td>* His10<\/sub>
* TwinStrepII
* S-tag
* CBP<\/td>		Ampicillin<\/td>pTT5<\/td><\/tr>
pCoofy58<\/td>VEGFss-TwinStrepII-3C<\/td>* His10<\/sub>
* TwinStrepII
* S-tag
* CBP<\/td>		Ampicillin<\/td>pTT5<\/td><\/tr>
pCoofy62<\/td>VEGFss-His6<\/sub>-SumoStar<\/td>* none
* His10<\/sub>
* TwinStrepII
* EPEA-tag<\/td>		Ampicillin<\/td>pTT5<\/td><\/tr><\/tbody><\/table>
* Multiple options for the C-terminal tag depending on the cloning strategy<\/figcaption><\/figure>\n\n<\/div>\n<\/div>\n\n\n
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\"diagram\"<\/a>
Vector map of the mammalian expression vector pXLG (David Hacker, EPFL Switzerland).<\/figcaption><\/figure>\n\n<\/div>\n<\/div>\n<\/div>\n\n\n\n
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pTT5 backbone:<\/h4>\n\n\n\n