The dominant expression of functional human lactoferrin in transgenic cloned goats using a hybrid lactoferrin expression construct
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- 作者：Huiqing Yu ; Jianquan Chen ; Wei Sun ; Siguo Liu ; Aimin Zhang ; Xujun Xu ; Xuebin Wang ; Zhuzi He ; Guohui Liu ; Guoxiang Cheng
- 关键词：Dominant expression ; Functional ; Human lactoferrin ; Cloned goat ; Goat milk
- 刊名：Journal of Biotechnology
- 出版年：2012
- 出版时间：31 October, 2012
- 年：2012
- 卷：161
- 期：3
- 页码：198-205
- 全文大小：922 K
文摘
Human Lactoferrin (hLF) is an iron-binding protein with multiple physiological functions. As the availability of natural hLF is limited, alternative means of producing this biopharmaceutical protein have been extensively studied. Here we report on the dominant expression of recombinant human lactoferrin (rhLF) in transgenic cloned goats using a novel optimised construct made by fusing a 3.3 kb hLF minigene to the regulatory elements of the ¦Â-casein gene. The transgenic goat produced more than 30 mg/ml rhLF in its milk, and rhLF expression was stable during the entire lactation cycle. The rhLF purification efficiency from whole goat milk is approximately 70 % , and its purity is above 98 % . Compared with natural hLF, the rhLF from transgenic goats has similar biological characteristics including molecular mass, N-terminal sequence, isoelectric point, immunoreactivity and digestive stability. More importantly, the purified rhLF showed specific anti-tumour activity in the mouse model of melanoma experimental metastasis. Therefore, our study shows that the large-scale production of functional rhLF in transgenic goat milk could be an economical and promising source of human therapeutic use in the future.