Expression of a lipocalin in prokaryote and eukaryote cells: quantification and structural characterization of recombinant bovine beta-lactoglobulin

Protein Expr Purif. 1999 Jun;16(1):70-5. doi: 10.1006/prep.1999.1055.

Abstract

In this paper we quantify and characterize the expression of recombinant beta-lactoglobulin (rBLG) in prokaryote and eukaryote cells. In Escherichia coli we used the pET26 vector, which permits the secretion of rBLG in periplasm. We studied the expression of rBLG in COS-7 cells and in vivo in mouse tibialis muscle. The expression of rBLG was measured by two immunoassays specific, respectively, for BLG in its native and denatured conformation. We observed that rBLG was essentially expressed in a denatured form in E. coli even in the periplasm, whereas rBLG in eukaryote cells was found in its native conformation.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • COS Cells
  • Cattle
  • Escherichia coli / genetics
  • Female
  • Gene Expression
  • Genetic Vectors
  • Lactoglobulins / biosynthesis
  • Lactoglobulins / chemistry*
  • Lactoglobulins / genetics*
  • Mice
  • Mice, Inbred BALB C
  • Muscle, Skeletal / metabolism
  • Protein Denaturation
  • Protein Folding
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Transfection

Substances

  • Lactoglobulins
  • Recombinant Proteins