Fan Baoliang, Li Ning
National Laboratory for Agro-biotechnology, China Agricultural University, Beijing, China, 100094.
Transgenic Res. 2009 Feb;18(1):99-112. doi: 10.1007/s11248-008-9221-4. Epub 2008 Oct 16.
We have designed and synthesized a gene encoding a fusion protein comprising Magainin2 and a carrier protein with the aim of screening for a suitable carrier protein expressing antibacterial peptides in the mammalian expression system. The antibacterial peptide Magainin2 was used as a model. Our results on mammalian cell expression showed that there was no exceptional splicing in the transfected CHO-s cells. Analysis of the transgenic mouse model revealed that the expression level of this fusion protein in one transgenic positive mouse was up to 10 g/l, which is close to the level of beta-casein in goat milk. The bioactivity analysis showed that the digested fusion protein had antibacterial activity. These results demonstrate that the synthetic gene of the carrier protein is suitable for expressing an antibacterial peptide in a mammalian cell system at high productivity and efficiency. Moreover, they demonstrate the potential for producing antibacterial peptides in a transgenic animal bioreactor on a large scale and inexpensively.
我们设计并合成了一个编码融合蛋白的基因,该融合蛋白由马盖宁2(Magainin2)和一种载体蛋白组成,目的是在哺乳动物表达系统中筛选一种合适的表达抗菌肽的载体蛋白。抗菌肽马盖宁2用作模型。我们在哺乳动物细胞表达方面的结果表明,转染的CHO-s细胞中没有异常剪接。对转基因小鼠模型的分析显示,一只转基因阳性小鼠中这种融合蛋白的表达水平高达10 g/l,这接近羊奶中β-酪蛋白的水平。生物活性分析表明,消化后的融合蛋白具有抗菌活性。这些结果表明,载体蛋白的合成基因适合在哺乳动物细胞系统中高效表达抗菌肽。此外,它们还证明了在转基因动物生物反应器中大规模且低成本生产抗菌肽的潜力。