Manual de expresion de proteinas

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Fifth Edition
The QIAexpressionist ™
A handbook for high-level expression and purification of 6xHis-tagged proteins March 2001

Trademarks Patented or patent-pending and/or registered or registration-pending trademarks of QIAGEN: QIAGEN®, QIAexpress®, QIAexpressionist ™, QIAquick®, Effectene™, HisSorb™, Ni-NTA, MinElute™, Penta·His™, PolyFect®, RGS·His™, SuperFect™, Tetra·His™. Hoffmann-LaRoche owns patents and patent applications pertaining to the application of Ni-NTA resin (Patent series: RAN 4100/63: USP 4.877.830, USP 5.047.513, EP 253 303 B1), and to 6xHis-coding vectors and His-labeled proteins (Patent series: USP 5.284.933, USP 5.310.663, EP 282 042 B1). All purification of recombinant proteins by Ni-NTA chromatography for commercial purposes, and the commercial use ofproteins so purified, require a license from Hoffmann-La Roche. ABTS is a registered trademark of Boehringer Mannheim GmbH. BaculoGold is a trademark of Becton, Dickinson and Company. BacVector is a registered trademark of Novagen, Inc. CDP-Star is a trademark of Tropix, Inc. Coomassie is a trademark of ICI Organics Inc. FPLC is a registered trademark of Pharmacia Biotech AB. High Five is a trademarkof Invitrogen Corporation Igepal is a registered trademark of Rhone-Poulenc, Inc. Macintosh is a registered trademark of Apple Computer, Inc. Microsoft is a registered trademark of Microsoft Corporation. Superflow is a trademark of Sterogene Bioseparations Inc. Triton is a registered trademark of Union Carbide Inc. Tween is a registered trademark of ICI Americas Inc. Registered names, trademarks,etc. used in this document, even when not specifically marked as such, are not to be considered unprotected by law.


The PCR process is covered by U.S. Patents 4,683,195 and 4,683,202 and foreign equivalents owned by Hoffmann-La Roche AG. © 2001 QIAGEN, all rights reserved.

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Contents
Kit Contents Storage Conditions Technical Assistance Product Use Limitations Product Warranty and Satisfaction Guarantee Safety Information Introduction and General Guidelines The QIAexpress System
QIAexpress pQE vectors QIAexpresspQE-TriSystem Vector for expression in E.coli, mammalian and insect cells Regulation of expression — pREP4 plasmid E. coli host strains The 6xHis tag Ni-NTA technology Ni-NTA chromatographic materials 7 8 9 9 9 10 11 15 15 16 16 17 18 18 19 21 21 22 22 23 23 23 23 24 24 28 29 29 35 36 3

Cloning
Choosing a QIAexpress construct Intended use of recombinant proteins and pQE vector choice Protein size...
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