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生产在乳汁中大量表达重组人溶菌酶的转基因克隆猪。

Production of transgenic-cloned pigs expressing large quantities of recombinant human lysozyme in milk.

作者信息

Lu Dan, Liu Shen, Shang Shengzhe, Wu Fangfang, Wen Xiao, Li Zhiyuan, Li Yan, Hu Xiaoxiang, Zhao Yaofeng, Li Qiuyan, Li Ning

机构信息

The State Key Laboratory for Agro-biotechnology, China Agricultural University, Beijing, China.

College of Animal Science and Technology, Yunnan Agricultural University, Kunming, China.

出版信息

PLoS One. 2015 May 8;10(5):e0123551. doi: 10.1371/journal.pone.0123551. eCollection 2015.

Abstract

Human lysozyme is a natural non-specific immune factor in human milk that plays an important role in the defense of breastfed infants against pathogen infection. Although lysozyme is abundant in human milk, there is only trace quantities in pig milk. Here, we successfully generated transgenic cloned pigs with the expression vector pBAC-hLF-hLZ-Neo and their first generation hybrids (F1). The highest concentration of recombinant human lysozyme (rhLZ) with in vitro bioactivity was 2759.6 ± 265.0 mg/L in the milk of F0 sows. Compared with wild-type milk, rhLZ milk inhibited growth of Escherichia coli K88 during the exponential growth phase. Moreover, rhLZ in milk from transgenic sows was directly absorbed by the intestine of piglets with no observable anaphylactic reaction. Our strategy may provide a powerful tool for large-scale production of this important human protein in pigs to improve resistance to pathogen infection.

摘要

人溶菌酶是母乳中的一种天然非特异性免疫因子,在保护母乳喂养的婴儿免受病原体感染方面发挥着重要作用。虽然溶菌酶在人乳中含量丰富,但在猪乳中仅微量存在。在此,我们成功利用表达载体pBAC-hLF-hLZ-Neo培育出了转基因克隆猪及其第一代杂交后代(F1)。F0代母猪乳汁中重组人溶菌酶(rhLZ)的最高浓度为2759.6±265.0毫克/升,且具有体外生物活性。与野生型乳汁相比,rhLZ乳汁在指数生长期抑制了大肠杆菌K88的生长。此外,转基因母猪乳汁中的rhLZ被仔猪肠道直接吸收,未观察到过敏反应。我们的策略可能为在猪中大规模生产这种重要的人类蛋白质以提高对病原体感染的抵抗力提供有力工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ffc/4425539/9f0b7c3fd32f/pone.0123551.g001.jpg

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