A uniform procedure for the purification of CDK7/CycH/MAT1, CDK8/CycC and CDK9/CycT1
<p>We have established a uniform procedure for the expression and purification of the cyclin-dependent kinases CDK7/CycH/MAT1, CDK8/CycC and CDK9/CycT1. We attach a His<sub>6</sub>-tag to one of the subunits of each complex and then co-express it together with the other subunits in...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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BMC
2004-01-01
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| Series: | Biological Procedures Online |
| Subjects: | |
| Online Access: | http://www.biologicalprocedures.com/bpo/arts/1/86/m86.htm |
| _version_ | 1811261026375565312 |
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| author | Pinhero Reena Liaw Peter Yankulov Krassimir |
| author_facet | Pinhero Reena Liaw Peter Yankulov Krassimir |
| author_sort | Pinhero Reena |
| collection | DOAJ |
| description | <p>We have established a uniform procedure for the expression and purification of the cyclin-dependent kinases CDK7/CycH/MAT1, CDK8/CycC and CDK9/CycT1. We attach a His<sub>6</sub>-tag to one of the subunits of each complex and then co-express it together with the other subunits in <it>Spodoptera frugiperda</it> insect cells. The CDK complexes are subsequently purified by Ni<sup>2+</sup>-NTA and Mono S chromatography. This approach generates large amounts of active recombinant kinases that are devoid of contaminating kinase activities. Importantly, the properties of these recombinant kinases are similar to their natural counterparts (Pinhero <it>et al</it>. 2004, Eur J Biochem 271:1004-14). Our protocol provides a novel systematic approach for the purification of these three (and possibly other) recombinant CDKs. |
| first_indexed | 2024-04-12T18:57:22Z |
| format | Article |
| id | doaj.art-1a9587194ff74339b131cfe5ce7a64c5 |
| institution | Directory Open Access Journal |
| issn | 1480-9222 |
| language | English |
| last_indexed | 2024-04-12T18:57:22Z |
| publishDate | 2004-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Biological Procedures Online |
| spelling | doaj.art-1a9587194ff74339b131cfe5ce7a64c52022-12-22T03:20:17ZengBMCBiological Procedures Online1480-92222004-01-016116317210.1251/bpo86A uniform procedure for the purification of CDK7/CycH/MAT1, CDK8/CycC and CDK9/CycT1Pinhero ReenaLiaw PeterYankulov Krassimir<p>We have established a uniform procedure for the expression and purification of the cyclin-dependent kinases CDK7/CycH/MAT1, CDK8/CycC and CDK9/CycT1. We attach a His<sub>6</sub>-tag to one of the subunits of each complex and then co-express it together with the other subunits in <it>Spodoptera frugiperda</it> insect cells. The CDK complexes are subsequently purified by Ni<sup>2+</sup>-NTA and Mono S chromatography. This approach generates large amounts of active recombinant kinases that are devoid of contaminating kinase activities. Importantly, the properties of these recombinant kinases are similar to their natural counterparts (Pinhero <it>et al</it>. 2004, Eur J Biochem 271:1004-14). Our protocol provides a novel systematic approach for the purification of these three (and possibly other) recombinant CDKs.http://www.biologicalprocedures.com/bpo/arts/1/86/m86.htmCyclin-Dependent KinasesBaculoviridaeIsolation and Purification |
| spellingShingle | Pinhero Reena Liaw Peter Yankulov Krassimir A uniform procedure for the purification of CDK7/CycH/MAT1, CDK8/CycC and CDK9/CycT1 Biological Procedures Online Cyclin-Dependent Kinases Baculoviridae Isolation and Purification |
| title | A uniform procedure for the purification of CDK7/CycH/MAT1, CDK8/CycC and CDK9/CycT1 |
| title_full | A uniform procedure for the purification of CDK7/CycH/MAT1, CDK8/CycC and CDK9/CycT1 |
| title_fullStr | A uniform procedure for the purification of CDK7/CycH/MAT1, CDK8/CycC and CDK9/CycT1 |
| title_full_unstemmed | A uniform procedure for the purification of CDK7/CycH/MAT1, CDK8/CycC and CDK9/CycT1 |
| title_short | A uniform procedure for the purification of CDK7/CycH/MAT1, CDK8/CycC and CDK9/CycT1 |
| title_sort | uniform procedure for the purification of cdk7 cych mat1 cdk8 cycc and cdk9 cyct1 |
| topic | Cyclin-Dependent Kinases Baculoviridae Isolation and Purification |
| url | http://www.biologicalprocedures.com/bpo/arts/1/86/m86.htm |
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