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Stable expression and rapid purification of Escherichia coli GroEL and GroES chaperonins.

Publication ,  Journal Article
Kamireddi, M; Eisenstein, E; Reddy, P
Published in: Protein expression and purification
October 1997

An Escherichia coli expression vector pRE (P. Reddy, A. Peterkofsky, and K. McKenney, 1989, Nucleic Acids Res. 17, 10473-10488), originally developed for the cloning and expression of lethal genes, was used for cloning and hyperexpression of GroEL and GroES genes. Regulated gene expression is achieved in the pRE vector under the tight control of the lambda PL promoter. Upon induction of the promoter, stable expression of GroEL to about 60% of the total cell protein was observed. Similarly, stable expression of GroES to about 40% of the total cell protein was achieved. GroES was found to be a heat-stable protein while GroEL was not. Both GroE chaperonins were purified in a single chromatographic step with a yield of about 100 mg GroEL and 25 mg GroES per liter of E. coli culture. GroE chaperonins purified by the protocols described here were active in the renaturation of urea-denatured rhodanese.

Duke Scholars

Published In

Protein expression and purification

ISSN

1046-5928

Publication Date

October 1997

Volume

11

Issue

1

Start / End Page

47 / 52

Location

united states

Related Subject Headings

  • Biochemistry & Molecular Biology
  • 0699 Other Biological Sciences
  • 0601 Biochemistry and Cell Biology
 

Citation

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Kamireddi, M., Eisenstein, E., & Reddy, P. (1997). Stable expression and rapid purification of Escherichia coli GroEL and GroES chaperonins. Protein Expression and Purification, 11(1), 47–52.
Kamireddi, M., E. Eisenstein, and P. Reddy. “Stable expression and rapid purification of Escherichia coli GroEL and GroES chaperonins.Protein Expression and Purification 11, no. 1 (October 1997): 47–52.
Kamireddi M, Eisenstein E, Reddy P. Stable expression and rapid purification of Escherichia coli GroEL and GroES chaperonins. Protein expression and purification. 1997 Oct;11(1):47–52.
Kamireddi, M., et al. “Stable expression and rapid purification of Escherichia coli GroEL and GroES chaperonins.Protein Expression and Purification, vol. 11, no. 1, Oct. 1997, pp. 47–52.
Kamireddi M, Eisenstein E, Reddy P. Stable expression and rapid purification of Escherichia coli GroEL and GroES chaperonins. Protein expression and purification. 1997 Oct;11(1):47–52.
Journal cover image

Published In

Protein expression and purification

ISSN

1046-5928

Publication Date

October 1997

Volume

11

Issue

1

Start / End Page

47 / 52

Location

united states

Related Subject Headings

  • Biochemistry & Molecular Biology
  • 0699 Other Biological Sciences
  • 0601 Biochemistry and Cell Biology