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Related Experiment Videos

Large-scale preparation of recombinant ovine prolactin and determination of its in vitro and in vivo activity.

H Leibovich1, N Raver, A Herman

  • 1Institute of Biochemistry, Food Science and Nutrition, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot 76100, Israel.

Protein Expression and Purification
|August 3, 2001
PubMed
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Researchers successfully produced recombinant ovine prolactin (oPRL) in E. coli. This biologically active oPRL stimulated cell proliferation and progesterone secretion, but did not affect milk production in ewes.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Endocrinology

Background:

  • Prolactin (PRL) is a key hormone regulating reproduction and lactation.
  • The specific biological functions of ovine prolactin (oPRL) require further investigation using purified recombinant forms.

Purpose of the Study:

  • To develop a method for producing biologically active recombinant ovine prolactin (oPRL).
  • To characterize the biological activity of the produced recombinant oPRL in vitro and in vivo.

Main Methods:

  • Recombinant bovine prolactin (PRL) gene was mutated to create a gene encoding ovine PRL (oPRL).
  • The oPRL gene was expressed in Escherichia coli, and the protein was purified.
  • Biological activity was assessed using Nb(2) cell proliferation, luciferase assays with oPRL receptors, and progesterone secretion assays.

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Main Results:

  • High yields of purified recombinant oPRL (over 98% monomeric) were obtained.
  • Recombinant oPRL demonstrated biological activity by stimulating Nb(2) cell proliferation, luciferase activity, and progesterone secretion.
  • Recombinant oPRL showed no galactopoietic effect in lactating ewes, unlike ovine growth hormone or placental lactogen.

Conclusions:

  • A functional recombinant oPRL was successfully produced and purified.
  • The recombinant oPRL exhibits specific biological activities relevant to reproduction.
  • This recombinant oPRL serves as a valuable tool for studying prolactin's role in sheep reproduction.