Doxycycline-regulated growth hormone gene expression system for swine

Genet Mol Res. 2012 Aug 29;11(3):2946-57. doi: 10.4238/2012.July.10.5.

Abstract

Although growth hormone gene transgenic animals are much larger than normal animals, they manifest differences that have adverse effects on survival due to overexpression of growth hormone. We developed a stable pig embryonic fibroblast cell line expressing pig growth hormone (pGH) using the Tet-On system, with which we can conditionally manipulate expression of pGH in vivo. Inducible expression of pGH was achieved by combining reserve Tet-controlled transcriptional activator and tetracycline-responsive element in a single plasmid. The mRNA expression of pGH was significantly increased compared to the non-induced group by about 10-fold. The controlled secretion of pGH induced by doxycycline was further tested in stably transfected cells. We conclude that inducible GH expression can be achieved in pig embryonic fibroblasts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Chromosomes, Mammalian / genetics
  • Doxycycline / pharmacology*
  • Drug Resistance / drug effects
  • Embryo, Mammalian / cytology
  • Enzyme-Linked Immunosorbent Assay
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Gene Expression Regulation / drug effects*
  • Genetic Vectors / genetics
  • Growth Hormone / genetics*
  • Growth Hormone / metabolism
  • Karyotyping
  • Lentivirus / drug effects
  • Lentivirus / genetics
  • Plasmids / metabolism
  • Pyrrolidinones / pharmacology
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sus scrofa / genetics*
  • Transgenes / genetics

Substances

  • Pyrrolidinones
  • blasticidin A
  • Growth Hormone
  • Doxycycline