Mukherjee Ayan, Garrels Wiebke, Talluri Thirumala R, Tiedemann Daniela, Bősze Zsuzsanna, Ivics Zoltán, Kues Wilfried A
Friedrich-Loeffler-Institut, Institut für Nutztiergenetik, Mariensee, Germany.
Medical School Hannover, Institute of Laboratory Animal Sciences, Hannover, Germany.
Sci Rep. 2016 Apr 18;6:24464. doi: 10.1038/srep24464.
We describe the expression of recombinant fluorescent proteins in the milk of two lines of transgenic pigs generated by Sleeping Beauty transposon-mediated genetic engineering. The Sleeping Beauty transposon consisted of an ubiquitously active CAGGS promoter driving a fluorophore cDNA, encoding either Venus or mCherry. Importantly, the fluorophore cDNAs did not encode for a signal peptide for the secretory pathway, and in previous studies of the transgenic animals a cytoplasmic localization of the fluorophore proteins was found. Unexpectedly, milk samples from lactating sows contained high levels of bioactive Venus or mCherry fluorophores. A detailed analysis suggested that exfoliated cells of the mammary epithelium carried the recombinant proteins passively into the milk. This is the first description of reporter fluorophore expression in the milk of livestock, and the findings may contribute to the development of an alternative concept for the production of bioactive recombinant proteins in the udder.
我们描述了通过睡美人转座子介导的基因工程产生的两系转基因猪的乳汁中重组荧光蛋白的表达情况。睡美人转座子由一个驱动荧光团cDNA的普遍活跃的CAGGS启动子组成，该荧光团cDNA编码金星（Venus）或mCherry。重要的是，荧光团cDNA不编码分泌途径的信号肽，并且在之前对转基因动物的研究中发现荧光团蛋白定位于细胞质中。出乎意料的是，来自泌乳母猪的乳汁样本中含有高水平的具有生物活性的金星或mCherry荧光团。详细分析表明，乳腺上皮的脱落细胞将重组蛋白被动地带入乳汁中。这是首次报道家畜乳汁中报告荧光团的表达情况，这些发现可能有助于开发一种在乳腺中生产具有生物活性的重组蛋白的替代概念。
