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In vitro evaluation of a mammary gland specific expression vector encoding recombinant human lysozyme for development of transgenic dairy goat embryos

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Abstract

A vector expressing human lysozyme (pBC1-hLYZ-GFP-Neo) was evaluated for gene and protein expression following liposome-mediated transformation of C-127 mouse mammary cancer cells. Cultures of G418-resistant clones were harvested 24–72 h after induction with prolactin, insulin and hydrocortisone. Target gene expression was analyzed by RT-PCR and Western blot and recombinant human lysozyme (rhLYZ) bacteriostatic activity was also evaluated. The hLYZ gene was correctly transcribed and translated in C-127 cells and hLYZ inhibited gram-positive bacterial growth, indicating the potential of this expression vector for development of a mammary gland bioreactor in goats. Guanzhong dairy goat skin fibroblasts transfected with pBC1-hLYZ-GFP-Neo were used to construct a goat embryo transgenically expressing rhLYZ by somatic nuclear transplantation with a blastocyst rate of 9.0 ± 2.8 %. These data establish the basis for cultivation of mastitis-resistant hLYZ transgenic goats.

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Acknowledgments

We are indebted to Mr. Qi Wang and Hui Feng for providing experimental material. We thank Ms. Ronghua Wu, Yani Bian and Mr. Xing Liu in experimental preparation. This study was supported by National Transgenenic New Species Breeding Program of China (No. 2011ZX08008-005; No. 2009ZX08008-007B).

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Authors and Affiliations

  1. Anhui Provincial Laboratory of Local Livestock and Poultry Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, Anhui, China

    Tao Gui, Meiling Zhang, Jianwen Chen, Yuanliang Zhang, Naru Zhou, Yu Zhang, Jia Tao, Liucai Sui, Yunsheng Li, Ya Liu, Xiaorong Zhang & Yunhai Zhang

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  1. Tao Gui
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  2. Meiling Zhang
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  3. Jianwen Chen
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  4. Yuanliang Zhang
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  5. Naru Zhou
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  6. Yu Zhang
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  7. Jia Tao
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  8. Liucai Sui
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  9. Yunsheng Li
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  10. Ya Liu
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  11. Xiaorong Zhang
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  12. Yunhai Zhang
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Corresponding author

Correspondence to Yunhai Zhang.

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Gui, T., Zhang, M., Chen, J. et al. In vitro evaluation of a mammary gland specific expression vector encoding recombinant human lysozyme for development of transgenic dairy goat embryos. Biotechnol Lett 34, 1445–1452 (2012). https://doi.org/10.1007/s10529-012-0930-7

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  • DOI: https://doi.org/10.1007/s10529-012-0930-7

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