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Expression of the Recombinant Porcine GH Gene In Vitro Using Tetracycline Inducible Expression System  

Kwon Mo Sun (Department of Physiology, Catholic University of Daegu School of Medicine)
Koo Bon Chul (Department of Physiology, Catholic University of Daegu School of Medicine)
Kim Teoan (Department of Physiology, Catholic University of Daegu School of Medicine)
Publication Information
Reproductive and Developmental Biology / v.29, no.1, 2005 , pp. 49-55 More about this Journal
Abstract
We cloned cDNA of the PGH(porcine growth hormone) gene and constructed retrovirus vector designed to express PGH gene under the regulation of CMV (cytomegalovirus) promoter. To maximize the expression, WPRE(woodchuck hepatitis virus posttranscriptional regulatory element) sequence was placed at the downstream of the PGH gene. After infection with recombinant viruses, approximately 1×10/sup 6/ PFF(porcine fetal fibroblast) cells released PGH protein into the media as much as 1,400 ng. In a subsequent experiment, a modifications of the retrovirus vector was made to express the PGH gene in a teracycline-inducible manner. In PFF cells carrying these viral vector sequences, addition of doxycycline to the media resulted in 2∼6 fold increase in PGH synthesis. In the modified retrovirus vectors, the WPRE sequence also played a role in boosting the effect of the tetracycline induction. This result indicates that our tetracycline-inducible expression system might be a promising candidate in alleviating the complicate physiological problems caused by constitutive expression of the exogenous genes in the transgenic animals.
Keywords
PGH; Tetracycline-inducible expression system; WPRE; PFF;
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