Korean Journal of Agricultural Science (농업과학연구)

Institute of Agricultural Science, CNU (충남대학교 농업과학연구소)
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Porcine tissue factor promoter induces specific expression of target gene in response to human serum in porcine aortic endothelial cells

  • No, Jin Gu (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Lee, Haesun (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Lee, Na Hyun (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Sun, Wu-Sheng (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Hwang, In-Sul (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Lee, Minguk (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Oh, Keon Bong (Animal Biotechnology Division, National Institute of Animal Science, RDA)
  • Received : 2021.04.13
  • Accepted : 2021.05.25
  • Published : 2021.06.01
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Abstract

Previously, we reported that tissue factor (Tf) was included in the list of differentially expressed genes as an upregulated gene in a rejected porcine heart after xenotransplantation into monkey. In this study, we analyzed that expression of Tf in aortic endothelial cells (pAEC) isolated from alpha 1,3-galactosyltransferase knockout pig in response to allogeneic porcine serum and xenogeneic human serum. The consequence was significant upregulation of Tf expression by responding to human serum compared with porcine serum. To analyze the function of Tf gene as a promoter, we constructed reporter vectors for expression of luciferase linked to 1,246 and 787 base pairs of porcine Tf (pTF1246 and pTF787), and 535 base pairs of human TF (hTF535) sequences including putative promoter regions and AP-1 biding site at the 5' end. The reporter vectors were transfected into pAEC including cytomegalovirus enhancer/chicken β-actin (CAG)-luciferase vector as a control. Luciferase assay showed that all of the promoters were insufficient to express luciferase compared with CAG promoter in basic culture conditions. Notably, pTF1246, pTF787, and hTF535 led to a significant increase of luciferase expression in response to human serum compared with porcine serum while no change of CAG. pTF1246 and pTF787 showed higher expression than hTF535. Taken together, our findings suggest that pTF1246 and pTF787 promoters could mediate target gene expression specifically at xenogeneic stress condition.

Keywords

Acknowledgement

본 연구는 농촌진흥청 연구사업 (PJ01422801)의 지원에 의해 이루어진 것임.

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