Kerekes Andrea, Hoffmann Orsolya Ivett, Iski Gergely, Lipták Nándor, Gócza Elen, Kues Wilfried A, Bősze Zsuzsanna, Hiripi László
Department of Animal Biotechnology, NARIC-Agricultural Biotechnology Institute, Gödöllö, Hungary.
Department of Biotechnology, Friedrich-Loeffler-Institut, Institut für Nutztiergenetik, Mariensee, Neustadt, Germany.
PLoS One. 2017 Oct 27;12(10):e0187214. doi: 10.1371/journal.pone.0187214. eCollection 2017.
Transgenic rabbits carrying mammary gland specific gene constructs are extensively used for excreting recombinant proteins into the milk. Here, we report refined phenotyping of previously generated Venus transposon-carrying transgenic rabbits with particular emphasis on the secretion of the reporter protein by exocrine glands, such as mammary, salivary, tear and seminal glands. The Sleeping Beauty (SB) transposon transgenic construct contains the Venus fluorophore cDNA, but without a signal peptide for the secretory pathway, driven by the ubiquitous CAGGS (CAG) promoter. Despite the absence of a signal peptide, the fluorophore protein was readily detected in milk, tear, saliva and seminal fluids. The expression pattern was verified by Western blot analysis. Mammary gland epithelial cells of SB-CAG-Venus transgenic lactating does also showed Venus-specific expression by tissue histology and fluorescence microscopy. In summary, the SB-CAG-Venus transgenic rabbits secrete the recombinant protein by different glands. This finding has relevance not only for the understanding of the biological function of exocrine glands, but also for the design of constructs for expression of recombinant proteins in dairy animals.
携带乳腺特异性基因构建体的转基因兔被广泛用于将重组蛋白分泌到乳汁中。在此，我们报告了对先前产生的携带金星转座子的转基因兔的精细表型分析，特别强调了外分泌腺（如乳腺、唾液腺、泪腺和精囊腺）对报告蛋白的分泌情况。睡美人（SB）转座子转基因构建体包含金星荧光蛋白cDNA，但没有分泌途径的信号肽，由普遍存在的CAGGS（CAG）启动子驱动。尽管没有信号肽，但在乳汁、眼泪、唾液和精液中很容易检测到荧光蛋白。通过蛋白质免疫印迹分析验证了表达模式。SB-CAG-金星转基因泌乳母兔的乳腺上皮细胞通过组织学和荧光显微镜检查也显示出金星特异性表达。总之，SB-CAG-金星转基因兔通过不同腺体分泌重组蛋白。这一发现不仅与理解外分泌腺的生物学功能有关，也与设计用于在奶牛中表达重组蛋白的构建体有关。
