High level expression of biologically active estrogen receptor in Saccharomyces cerevisiae.

Published

Journal Article

Biochemical over-expression of the human estrogen receptor was achieved using a Saccharomyces cerevisiae expression system. The receptor was produced as a novel ubiquitin fusion protein. This fusion protein is short lived in the cell and is processed to produce unfused receptor shortly after folding. Conventional high copy expression plasmids produced receptor to about 0.04% of the total soluble protein. By incorporating a defective leu2 allele into these vectors, an additional 5-fold increase in receptor production was obtained. The recombinant receptor was undergraded, soluble and biologically active. Conventional methods of disrupting cells using glass beads had a detrimental effect on the ability of the receptor to bind hormone. Enzymatic digestion of the cell wall followed by hypotonic shock liberates the receptor that quantitatively binds estrogen.

Full Text

Duke Authors

Cited Authors

  • McDonnell, DP; Nawaz, Z; Densmore, C; Weigel, NL; Pham, TA; Clark, JH; O'Malley, BW

Published Date

  • September 1, 1991

Published In

Volume / Issue

  • 39 / 3

Start / End Page

  • 291 - 297

PubMed ID

  • 1911419

Pubmed Central ID

  • 1911419

International Standard Serial Number (ISSN)

  • 0960-0760

Language

  • eng

Conference Location

  • England