{"id":1815,"date":"2021-06-05T16:48:06","date_gmt":"2021-06-05T16:48:06","guid":{"rendered":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/?page_id=1815"},"modified":"2022-01-25T16:57:37","modified_gmt":"2022-01-25T16:57:37","slug":"insect-cell-expression-vectors","status":"publish","type":"page","link":"https:\/\/www.embl.org\/groups\/protein-expression-purification\/services\/strategy-and-construct-design\/insect-cell-expression-vectors\/","title":{"rendered":"Insect cell expression vectors"},"content":{"rendered":"\n
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Overview of our insect cell expression vectors<\/a><\/p>\n\n\n\n

Please note that only the vectors indicated in yellow can be shared with external users via the EMBL MTA.<\/p>\n\n\n\n

We also have a collection of insect cell 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

Insect cell pCOOFY expression vectors<\/h3>\n\n\n\n
Name<\/th>N-terminal tag<\/th>C-terminal tag<\/th>Antibiotic resistance<\/th><\/tr><\/thead>
pCoofy27<\/td>His7<\/sub>-3C<\/td>X<\/td>Ampicillin Gentamycin<\/td><\/tr>
pCoofy28<\/td>His6<\/sub>-GST-3C<\/td>X<\/td>Ampicillin Gentamycin<\/td><\/tr>
pCoofy29<\/td>His6<\/sub>-MBP-3C<\/td>X<\/td>Ampicillin Gentamycin<\/td><\/tr>
pCoofy41<\/td>3C (not expressed)<\/td>* His10<\/sub>
* TwinStrepII
* S-tag
* CBP<\/td>		Ampicillin Gentamycin<\/td><\/tr>
pCoofy51<\/td>TwinStrepII-3C<\/td>X<\/td>Ampicillin Gentamycin<\/td><\/tr>
pCoofy59<\/td>TrxA-His6<\/sub>-3C<\/td>X<\/td>Ampicillin Gentamycin<\/td><\/tr>
pCoofy60<\/td>His6<\/sub>-eGFP-3C<\/td>X<\/td>Ampicillin Gentamycin<\/td><\/tr>
pCoofy63<\/td>His6<\/sub>-SumoStar (Smt3 R64T R71E)<\/td>* none
* His10<\/sub>
* TwinStrepII
* EPEA-tag<\/td>		Ampicillin Gentamycin<\/td><\/tr>
pCoofy64<\/td>GP67ss-His6<\/sub>-SumoStar (Smt3 R64T R71E)<\/td>* none
* His10<\/sub>
* TwinStrepII
* EPEA-tag<\/td>		Ampicillin Gentamycin<\/td><\/tr><\/tbody><\/table>
* Multiple options for the C-terminal tag depending on the cloning strategy<\/figcaption><\/figure>\n\n\n\n

For the expression of protein complexes<\/strong> in insect cells, we have both the MultiBac<\/strong> and biGBac<\/strong> vectors available.<\/p>\n\n\n\n

More information about how to use the MultiBac system can be found on the websites of Geneva Biotech<\/a> and the University of Bristol<\/a>.<\/p>\n\n\n\n

A detailed description of the biGBac system can be found in the following references:<\/p>\n\n\nWeissmann F.,\u00a0Petzold G.,\u00a0VanderLinden R.,\u00a0Huis in ‘t Veld P.J.,\u00a0Brown N.G.,\u00a0Lampert F., Westermann S., Stark H., Schulman B.A. and\u00a0 Peters J.-M. (2016) biGBac enables rapid gene assembly for the expression of large multisubunit protein complexes. PNAS<\/em>. 113(19)<\/strong>: E2564-2569<\/blockquote>\n\n\nWeissmann F. and Peters J.-M. (2018) Expressing Multi-subunit Complexes Using biGBac. Methods Mol Biol.<\/em> 1764<\/strong>: 329-343<\/blockquote>\n\n<\/div>\n<\/div>\n\n\n

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