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Related Experiment Videos

[Murine beta-casein gene sequences direct a human tissue plasminogen activator variant expressed in the milk of

Fu-Yu Lin1, Xuan Cheng, Hong-Xing Chen

  • 1Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.

Yi Chuan Xue Bao = Acta Genetica Sinica
|April 16, 2003
PubMed
Summary

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Researchers successfully used murine beta-casein gene regulatory elements to express human tissue plasminogen activator (t-PA) variant in the milk of transgenic mice. This demonstrates a foundational method for creating mammary gland bioreactor models.

Area of Science:

  • Biotechnology
  • Genetics
  • Molecular Biology

Background:

  • Developing transgenic animal models for therapeutic protein production is crucial.
  • Mammary gland expression systems offer high yields for recombinant proteins.
  • The murine beta-casein gene promoter is a well-characterized regulatory element for milk-specific expression.

Purpose of the Study:

  • To assess the efficacy of murine beta-casein gene regulatory elements in directing exogenous gene expression in transgenic mouse milk.
  • To construct and validate a transgenic mouse model for human tissue plasminogen activator (t-PA) variant production.

Main Methods:

  • Construction of a mammary gland expression vector containing a fusion gene with human t-PA variant cDNA under murine beta-casein regulatory control.
  • Microinjection of the fusion gene into fertilized mouse eggs and transfer to surrogate mothers.

Related Experiment Videos

  • Identification of transgenic mice using PCR and Southern blot analysis.
  • Quantification of human t-PA variant expression in the milk of positive transgenic mice.

    • Main Results:

    • Twelve out of 42 newborn mice were confirmed as positive transgenic individuals.
    • Human t-PA variant was successfully expressed in the milk of 7 transgenic mice.
    • The highest expression level of human t-PA variant reached 3.6593 micrograms/ml.

    Conclusions:

    • Murine beta-casein gene regulatory elements effectively direct the expression of the human t-PA variant gene in the milk of transgenic mice.
    • This study provides a strong foundation for constructing beta-casein knock-in mice mammary gland bioreactor models.
    • The developed model shows potential for efficient production of therapeutic proteins in milk.