Manual de expresion de proteinas

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...