Abstract

Doxycycline-regulated growth hormone gene expression system for swine

Author(s): B.C. Jiang D.B. Yu L.J. Wang F.L. Dong H.A. Kaleri X.G. Wang N. Ally J. Li H.L. Liu

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. 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.

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