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The High-Level Expression of Human Tissue Plasminogen Activator in the Milk of Transgenic Mice with Hybrid Gene Locus Strategy

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Abstract

Transgene expression for the mammary gland bioreactor aimed at producing recombinant proteins requires optimized expression vector construction. Previously we presented a hybrid gene locus strategy, which was originally tested with human lactoferrin (hLF) as target transgene, and an extremely high-level expression of rhLF ever been achieved as to 29.8 g/l in mice milk. Here to demonstrate the broad application of this strategy, another 38.4 kb mWAP-htPA hybrid gene locus was constructed, in which the 3-kb genomic coding sequence in the 24-kb mouse whey acidic protein (mWAP) gene locus was substituted by the 17.4-kb genomic coding sequence of human tissue plasminogen activator (htPA), exactly from the start codon to the end codon. Corresponding five transgenic mice lines were generated and the highest expression level of rhtPA in the milk attained as to 3.3 g/l. Our strategy will provide a universal way for the large-scale production of pharmaceutical proteins in the mammary gland of transgenic animals.

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Acknowledgments

This research was supported by the fund 2009ZX10004-402 from National Major Science and Technology Program for Infectious Disease.

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Authors and Affiliations

  1. Cell Engineering Department, Beijing Institute of Biotechnology, No. 20 Dongdajie Street, Beijing, 100071, China

    Yanrong Zhou, Yanli Lin, Xiaojie Wu, Fuyin Xiong, Yuemeng Lv, Tao Zheng, Peitang Huang & Hongxing Chen

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  1. Yanrong Zhou
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  2. Yanli Lin
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  3. Xiaojie Wu
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  4. Fuyin Xiong
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  5. Yuemeng Lv
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  6. Tao Zheng
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  7. Peitang Huang
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  8. Hongxing Chen
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Correspondence to Hongxing Chen.

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The authors Yanrong Zhou, Yanli Lin, and Xiaojie Wu contributed equally to this work.

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Zhou, Y., Lin, Y., Wu, X. et al. The High-Level Expression of Human Tissue Plasminogen Activator in the Milk of Transgenic Mice with Hybrid Gene Locus Strategy. Mol Biotechnol 50, 137–144 (2012). https://doi.org/10.1007/s12033-011-9428-0

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