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The co-promoting expression analysis of the tPA/GH double transgenic goat mammary epithelial cells and thrombolytic activity of the tPA in-vitro
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  • Shaozheng Song,
  • Yaoling Yaoling Luo,
  • Zhaoxia Liu,
  • Dan Li,
  • Junsong Junsong Ye,
  • Zhengyi He
Shaozheng Song
Taihu University of Wuxi
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Yaoling Yaoling Luo
The First Affiliated Hospital of Gannan Medical University
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Zhaoxia Liu
The First Affiliated Hospital of Gannan Medical University
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Dan Li
Taihu University of Wuxi
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Junsong Junsong Ye
The First Affiliated Hospital of Gannan Medical University

Corresponding Author:[email protected]

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Zhengyi He
The First Affiliated Hospital of Gannan Medical University
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Abstract

The human tissue-plasminogen activator (tPA) is a thrombolytic drug widely used in the treatment of stroke, pulmonary thrombosis, acute myocardial infarction, and other thrombotic diseases. The double gene co-integration into the organisms and cells can produce a synergistic effect, which can improve the expression level of the target gene. We selected the mammary gland-specific expression vectors BLC14/tPA and BLC14/GH with the β-lactoglobulin gene as a regulatory sequence in our previous research. The tPA and GH were co-transfected into goat mammary epithelial cells by electro-transfection. The results showed that the expression level of the tPA in single-gene cells was 8.0-64.0 μg/mL, while in double-gene cells was 200-7200 μg/mL, which was significantly higher than that in single-gene cells. In addition, the tPA also had a relatively strong in-vitro thrombolytic activity function determined by FAPA. The results showed that the goat mammary epithelial cell lines with the tPA/GH gene integration were successfully obtained by electro-transfection and it was proved that the expression level of the tPA in the double gene integration cell lines with the tPA/GH genes integration was significantly increased, which provided a new way for the preparation of highly expressed transgenic goat and other animals by somatic cell nuclear transfer, and also laid a foundation for the efficient production of pharmaceutical proteins in transgenic animal mammary gland reactors in the future.