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Transgenic rabbit producing human growth hormone in milk
Daniel Lipiński1, Jolanta Jura, Robert Kalak
1Department of Biochemistry and Biotechnology, August Cieszkowski Agricultural University of Poznań, ul. Wołyńska 35, 60-637 Poznań, Poland.
Journal of Applied Genetics
|May 30, 2003
Summary
Researchers developed a novel gene construct for producing human growth hormone (hGH) in mammal mammary glands. This genetically engineered hGH can be efficiently purified from milk using affinity chromatography.
Area of Science:
- Biotechnology
- Genetics
- Molecular Biology
Background:
- Mammary gland expression systems are crucial for producing recombinant proteins.
- Efficient purification methods are needed for therapeutic proteins like human growth hormone (hGH).
Purpose of the Study:
- To create a gene construct for expressing human growth hormone (hGH) in the mammary glands of mammals.
- To develop a method for easy purification of biologically active hGH from milk.
Main Methods:
- A gene construct, WAP(6xHisThr):hGH, was engineered with a rat whey acidic protein (WAP) promoter and modifications for purification.
- The construct was introduced into rabbit oocytes via microinjection.
- Transgenic rabbits were generated, and hGH expression was analyzed in milk from F1 generation females.
Main Results:
- The WAP(6xHisThr):hGH gene construct was successfully created and integrated into the rabbit genome (chromosome 7).
- Human growth hormone (hGH) was detected in the milk of lactating F1 generation rabbits.
- The engineered hGH, with a six-histidine tag and thrombin cleavage site, was amenable to purification via affinity chromatography and thrombin cleavage.
Conclusions:
- Mammary gland-specific expression of human growth hormone (hGH) in transgenic rabbits is feasible using the WAP promoter.
- The developed gene construct allows for the efficient production and purification of active hGH from milk, offering a potential biopharmaceutical production strategy.