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High-Level Expression, Purification, and Renaturation of Recombinant Murine Interleukin-2 from Escherichia coli
- Source :
- Protein Expression and Purification. 4:240-246
- Publication Year :
- 1993
- Publisher :
- Elsevier BV, 1993.
-
Abstract
- A murine interleukin-2 (mIL-2)-encoding cDNA, isolated from a stimulated EL4 mRNA library, was used to construct several expression plasmids directing synthesis of the mature protein in Escherichia coli. The expression was under control of either the PTrp or the PL promoter. Using these systems, a high-level expression of between 10 and 35% of the total cellular protein was obtained. The mIL-2 protein, present as insoluble inclusion bodies, could be solubilized in a chaotropic mixture and was partially purified by preparative gel filtration under denaturing conditions. After renaturation, the protein was further purified to homogeneity by anion-exchange chromatography. Depending on the fermentation, induction, and renaturation conditions, the yield ranged between 0.35 and 1 mg of purified mIL-2/g wet cells. The specific biological activity was about 10(7) units/mg and the endotoxin content < 4 ng/mg pure recombinant protein.
- Subjects :
- Protein Denaturation
Protein Folding
Molecular Sequence Data
Size-exclusion chromatography
Gene Expression
Biology
medicine.disease_cause
Inclusion bodies
law.invention
Mice
law
Complementary DNA
Gene expression
Escherichia coli
medicine
Animals
Amino Acid Sequence
Inclusion Bodies
Messenger RNA
Base Sequence
Molecular biology
Recombinant Proteins
Chaotropic agent
Solubility
Biochemistry
Recombinant DNA
Interleukin-2
Plasmids
Biotechnology
Subjects
Details
- ISSN :
- 10465928
- Volume :
- 4
- Database :
- OpenAIRE
- Journal :
- Protein Expression and Purification
- Accession number :
- edsair.doi.dedup.....b65acffb4ad32f34978f243ac2f654a3