Human nerve growth factor beta (hNGF-β):: mammary gland specific expression and production in transgenic rabbits
Human nerve growth factor beta (hNGF-β):: mammary gland specific expression and production in transgenic rabbits
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DOI：
10.1016/s0014-5793(98)01728-1
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发表时间：
1999-02-05
期刊：
影响因子：
3.5
通讯作者：
Brem, G
中科院分区：
文献类型：
--
作者：
Coulibaly, S;Besenfelder, U;Brem, G
产生携带编码人神经生长因子β（hNGF-β）cDNA的基因构建体的转基因兔。hNGF-beta mRNA的表达仅限于哺乳期兔的乳腺。WesternBlot分析显示转基因动物乳汁中含有13.2kDa的多肽，通过两步色谱法从乳汁中纯化hNGF-β，纯化的hNGF-β的电喷雾质谱分析描绘了每个亚基的分子量为13261 Da。使用PC 12 W2细胞和来自鸡胚的背根神经节神经元培养物测试hNGF-β的生物活性。与商业重组hNGF-β相比，来自转基因动物的粗脱脂乳和纯化的hNGF-β表现出完全的生物活性。(C)1999年欧洲生物化学学会联合会。
Transgenic rabbits carrying gene constructs encoding human nerve growth factor beta (hNGF-beta) cDNA were generated. Expression of hNGF-beta mRNA was restricted to the mammary gland of lactating rabbits. Western Blot analysis revealed a polypeptide of 13.2 kDa in the milk of transgenic animals, hNGF-beta was purified from the milk by a two-step chromatographic procedure, Electrospray mass spectroscopy analysis of purified hNGF-beta depicted a molecular weight of 13 261 Da per subunit. The biological activity of the hNGF-beta was tested using PC12W2 cells and cultures of dorsal root ganglion neurons from chicken embryos. Crude defatted milk from transgenic animals and purified hNGF-beta demonstrated full biological activity when compared to commercial recombinant hNGF-beta. (C) 1999 Federation of European Biochemical Societies.