Purification of recombinantly expressed human cluster determinant 4 cytoplasmic domain
| Autor: | Dieter Willbold, Gesa Jonas, Andrea Preusser |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Clinical Biochemistry
Peptide medicine.disease_cause Biochemistry Chromatography Affinity Analytical Chemistry law.invention Gel permeation chromatography Plasmid law medicine Humans A-DNA Escherichia coli DNA Primers chemistry.chemical_classification Chromatography Expression vector Membrane Glycoproteins Base Sequence Cell Biology General Medicine Recombinant Proteins chemistry Cytoplasm Recombinant DNA Chromatography Gel Electrophoresis Polyacrylamide Gel |
| Zdroj: | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences. 786(1-2) |
| ISSN: | 1570-0232 |
| Popis: | A DNA fragment coding for the human CD4 cytoplasmic domain (residues 394-433) was cloned into the pET15b expression vector. The resulting plasmid was used for synthesis of the polyhistidine-tagged 5.10(3) M(r) CD4 peptide in Escherichia coli BL21(DE3)Star. The CD4 cytoplasmic domain was purified under denaturing and reducing conditions by a two-step procedure using immobilized metal affinity chromatography and gel permeation chromatography. The purified CD4 cytoplasmic domain is soluble and functional without any specific refolding steps. The yield of the described purification procedure was approximately 5 mg peptide per liter culture volume. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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