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Doxycycline-regulated growth hormone gene expression system for swine
1Department of Animal Breeding & Genetics, College of Animal Science & Technology, Nanjing Agricultural University, Nanjing, China.
Genetics and Molecular Research : GMR
|August 8, 2012
Summary
Researchers created a pig embryonic fibroblast cell line for controlled pig growth hormone (pGH) expression. This Tet-On system allows inducible pGH manipulation in vivo, aiding future research.
Area of Science:
- Biotechnology
- Animal Science
- Molecular Biology
Background:
- Transgenic animals overexpressing growth hormone exhibit enhanced size but often face adverse survival effects.
- Controlling gene expression is crucial for mitigating negative impacts of growth hormone overexpression.
Purpose of the Study:
- To develop a stable pig embryonic fibroblast cell line with conditionally inducible pig growth hormone (pGH) expression.
- To establish a system for precise control over pGH levels in vitro and potentially in vivo.
Main Methods:
- Utilized the Tet-On inducible gene expression system.
- Constructed a single plasmid containing the Tet-controlled transcriptional activator and tetracycline-responsive element.
- Developed a stable pig embryonic fibroblast cell line transfected with the pGH construct.
Main Results:
- Achieved significantly increased mRNA expression of pGH (approximately 10-fold) in the induced group compared to the non-induced control.
- Demonstrated controlled secretion of pGH in stably transfected cells upon doxycycline induction.
Conclusions:
- Successfully established a method for inducible growth hormone expression in pig embryonic fibroblasts.
- The Tet-On system provides a viable strategy for conditional pGH manipulation, offering potential for safer transgenic applications.
