Recombinant human CIS2 (SOCS2) protein

Subcloning, expression, purification, and characterization

Eva Biener, Sarah Maurice, Yael Sandowski, Yael Cohen, Eugene E. Gusakowsky, Robert Hooghe, Akihiko Yoshimura, Oded Livnah, Arieh Gertler

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The 1x myc-tagged cDNA encoding for human CIS2 protein was subcloned into a pET-29a+ vector in order to express and produce a recombinant S-peptide tagged and 1x myc-tagged protein in Escherichia coli BL21(DE3). The constitutively expressed protein was isolated from inclusion bodies by a simple solubilization-renaturation procedure and purified by anion-exchange chromatography on Q-Sepharose. The recombinant form was found to be pure and monomeric as judged by both SDS-PAGE and gel-filtration chromatography and its biological activity was proven by its ability to bind to the tyrosine-phosphorylated cytosolic fragment of human growth hormone receptor fused to glutathione-S-transferase. Recombinant CIS2 was compared by biochemical, immunological, and molecular methods to the CIS2 protein expressed in eukaryotic cells. This report describes the first substantial production of biologically active recombinant human CIS2.

Original languageEnglish
Pages (from-to)305-312
Number of pages8
JournalProtein Expression and Purification
Volume25
Issue number2
DOIs
Publication statusPublished - 2002
Externally publishedYes

Fingerprint

Purification
Chromatography
Escherichia coli Proteins
Inclusion Bodies
Eukaryotic Cells
Glutathione Transferase
Sepharose
Gel Chromatography
Anions
Tyrosine
Polyacrylamide Gel Electrophoresis
Proteins
Complementary DNA
Bioactivity
Escherichia coli
Peptides
Gels
human SOCS2 protein
delta-hGHR

Keywords

  • CIS2
  • Growth hormone receptor
  • Recombinant proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Biener, E., Maurice, S., Sandowski, Y., Cohen, Y., Gusakowsky, E. E., Hooghe, R., ... Gertler, A. (2002). Recombinant human CIS2 (SOCS2) protein: Subcloning, expression, purification, and characterization. Protein Expression and Purification, 25(2), 305-312. https://doi.org/10.1016/S1046-5928(02)00013-X

Recombinant human CIS2 (SOCS2) protein : Subcloning, expression, purification, and characterization. / Biener, Eva; Maurice, Sarah; Sandowski, Yael; Cohen, Yael; Gusakowsky, Eugene E.; Hooghe, Robert; Yoshimura, Akihiko; Livnah, Oded; Gertler, Arieh.

In: Protein Expression and Purification, Vol. 25, No. 2, 2002, p. 305-312.

Research output: Contribution to journalArticle

Biener, E, Maurice, S, Sandowski, Y, Cohen, Y, Gusakowsky, EE, Hooghe, R, Yoshimura, A, Livnah, O & Gertler, A 2002, 'Recombinant human CIS2 (SOCS2) protein: Subcloning, expression, purification, and characterization', Protein Expression and Purification, vol. 25, no. 2, pp. 305-312. https://doi.org/10.1016/S1046-5928(02)00013-X
Biener, Eva ; Maurice, Sarah ; Sandowski, Yael ; Cohen, Yael ; Gusakowsky, Eugene E. ; Hooghe, Robert ; Yoshimura, Akihiko ; Livnah, Oded ; Gertler, Arieh. / Recombinant human CIS2 (SOCS2) protein : Subcloning, expression, purification, and characterization. In: Protein Expression and Purification. 2002 ; Vol. 25, No. 2. pp. 305-312.
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