Abstract
Transgenesis has been used for expressing human lysozyme (hLZ) in the milk of livestock to improve their disease resistance. Here we describe a human lactoferrin (hLF) BAC as a candidate vector for high-level expression of hLZ in the milk of transgenic mice. Using recombineering, hLF genomic DNA in the hLF BAC was replaced by the hLZ gene (from the ATG start codon to the TAA stop codon), and flanking regions of the hLF gene (a 90-kb 5′ and a 30-kb 3′) were used as transcriptional control elements for hLZ expression. When this construct was used to generate transgenic mice, rhLZ was highly expressed in the milk of four transgenic mouse lines (1.20–1.76 g/L), was expressed at a lower level in one additional line (0.21 g/L). rhLZ from the milk of these transgenic mice exhibited the same antibacterial activity as native hLZ. Our results suggest a potential approach for producing large amounts of hLZ in the milk of livestock.
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Acknowledgments
We would like to thank Drs. Yaofeng Zhao, Sen Wu, Jia Tong, Guangbin Zhou, Jianwu Wang, Tian Yu, Jin He and Jianxiang Xu for revising the manuscript, Hongxing Chen for his discussions on BAC modification, and Min Zhang, Rui Fang, Mingjun Bi, Zubin Cao, Zhisheng Chen and Junna He for excellent technical assistance. This work was supported by “863” High-Tech Research Development (Project Grant No. 2009AA10Z110 and 2010AA10A103).
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Liu, S., Li, X., Lu, D. et al. High-level expression of bioactive recombinant human lysozyme in the milk of transgenic mice using a modified human lactoferrin BAC. Transgenic Res 21, 407–414 (2012). https://doi.org/10.1007/s11248-011-9536-4
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DOI: https://doi.org/10.1007/s11248-011-9536-4