Coulibaly S, Besenfelder U, Fleischmann M, Zinovieva N, Grossmann A, Wozny M, Bartke I, Tögel M, Müller M, Brem G
Ludwig Boltzmann Institute for Immuno-, Cyto- and Molecular Genetic Research, Vienna, Austria.
FEBS Lett. 1999 Feb 5;444(1):111-6. doi: 10.1016/s0014-5793(98)01728-1.
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.
生成了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-β mRNA的表达局限于泌乳兔的乳腺。蛋白质免疫印迹分析显示转基因动物乳汁中有一条13.2 kDa的多肽。通过两步色谱法从乳汁中纯化出hNGF-β。对纯化后的hNGF-β进行电喷雾质谱分析，结果表明每个亚基的分子量为13261 Da。使用PC12W2细胞和鸡胚背根神经节神经元培养物对hNGF-β的生物活性进行了测试。与市售重组hNGF-β相比，转基因动物的粗脱脂乳和纯化后的hNGF-β均表现出完全的生物活性。
