High level expression of a truncated chicken progesterone receptor in Escherichia coli.

Journal Article (Journal Article)

Using a novel Escherichia coli system we have successfully overexpressed a region of the chicken progesterone receptor which encodes both the DNA- and hormone-binding domains. The expression system produces the truncated receptor fragment as an in-frame fusion with ubiquitin. This strategy greatly enhances both the solubility and stability of fusion proteins expressed in E. coli. Synthesis has been further improved by induction of the lambda PL promoter with nalidixic acid at low growth temperatures (less than or equal to 30 degrees C) rather than use of conventional heat induction protocols. We can produce 10 mg of receptor fragment/liter of cells using this system, and we estimate that at least 0.3 mg of this receptor material is biologically active, as assessed by DNA-binding and hormone-binding assays. Receptor produced in this manner is almost indistinguishable from authentic oviduct progesterone receptor using the criteria of hormone-binding specificity and affinity and binding to a progesterone response element. This expression system offers a cheap convenient method for the production of mg amounts of biologically active derivatives of progesterone receptor for biochemical studies.

Full Text

Duke Authors

Cited Authors

  • Power, RF; Conneely, OM; McDonnell, DP; Clark, JH; Butt, TR; Schrader, WT; O'Malley, BW

Published Date

  • January 25, 1990

Published In

Volume / Issue

  • 265 / 3

Start / End Page

  • 1419 - 1424

PubMed ID

  • 2153132

International Standard Serial Number (ISSN)

  • 0021-9258


  • eng

Conference Location

  • United States