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High-level expressing YAC vector for transgenic animal bioreactors.

Y Fujiwara1, M Miwa, R Takahashi

  • 1YS New Technology Institute, Inc., Ishibashi-machi, Shimotsuga-gun, Tochigi, Japan.

Molecular Reproduction and Development
|March 26, 1999
PubMed
Summary
This summary is machine-generated.

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This study developed transgenic rats using yeast artificial chromosome (YAC) vectors to produce human growth hormone (hGH) in milk. The YAC vector enabled high-level, tissue-specific hGH expression, overcoming position effects in transgenic animal production.

Area of Science:

  • Biotechnology
  • Genetics
  • Animal Science

Background:

  • Position effect is a significant challenge in creating transgenic animals for bioreactor applications.
  • Efficient gene expression in mammary glands is crucial for producing therapeutic proteins.

Purpose of the Study:

  • To assess the utility of a 210-kb human alpha-lactalbumin YAC vector for producing transgenic rats.
  • To evaluate the efficiency and expression levels of human growth hormone (hGH) in the milk of these transgenic rats.

Main Methods:

  • Homologous recombination was used to insert the hGH gene into YAC vectors.
  • YAC DNA was microinjected into rat embryos to generate transgenic rats.
  • hGH expression and tissue specificity were analyzed in the milk and mRNA of transgenic rats.

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Main Results:

  • Transgenic rats were successfully produced with YAC vector DNA, showing similar efficiency to plasmid constructs.
  • All analyzed transgenic rats carried one copy of the transgene and secreted high levels of hGH (0.25-8.9 mg/ml) in their milk.
  • hGH mRNA exhibited tissue specificity mirroring the endogenous rat alpha-lactalbumin gene.

Conclusions:

  • The 210-kb human alpha-lactalbumin YAC vector is effective for high-level, tissue-specific expression of foreign genes in transgenic animal milk.
  • This YAC vector system offers a promising solution for overcoming position effects in mammary gland bioreactor production.
  • The findings support the use of YAC vectors for efficient production of therapeutic proteins in transgenic animals.