College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China.
Transgenic Res. 2013 Aug;22(4):805-12. doi: 10.1007/s11248-012-9682-3. Epub 2013 Jan 18.
The feasibility of using the pig parotid secretory protein promoter to drive the β-glucanase transgene expression in mouse parotid glands was examined in this study. The parotid gland-specific vector expressing β-glucanase gene (GLU, from Paenibacillus polymyxa CP7) was constructed. Transgenic mice were produced by the pronuclear microinjection. Both PCR and Southern blot analysis showed that the mice carried the β-glucanase gene and the β-glucanase gene could be stably inherited. Furthermore, RT-PCR and northern blot analysis indicated that it was specifically expressed in the parotid. The β-glucanase activity in the saliva was found to be 0.18 U/mL. After feeding a diet containing 2 % β-glucan, the average daily gain of transgenic was significantly higher than non-transgenic mice. The crude protein and crude fat concentration in faeces of transgenic mice were significantly reduced compared with that of the non-transgenic mice. These results suggest that the successful expression of foreign β-glucanase in the animal parotid would offer a promising biological approach to reduce the anti-nutritional effect of β-glucans in feed.
本研究探讨了利用猪腮腺分泌蛋白启动子驱动β-葡聚糖酶转基因在小鼠腮腺中表达的可行性。构建了表达β-葡聚糖酶基因(来自多粘类芽孢杆菌 CP7 的 GLU)的腮腺特异性载体。通过原核显微注射生产转基因小鼠。PCR 和 Southern blot 分析均表明,小鼠携带β-葡聚糖酶基因,且β-葡聚糖酶基因可稳定遗传。此外,RT-PCR 和 northern blot 分析表明其在腮腺中特异性表达。唾液中的β-葡聚糖酶活性为 0.18 U/mL。喂食含有 2%β-葡聚糖的饲料后,转基因小鼠的平均日增重明显高于非转基因小鼠。与非转基因小鼠相比,转基因小鼠粪便中的粗蛋白和粗脂肪浓度明显降低。这些结果表明,外源β-葡聚糖酶在动物腮腺中的成功表达为降低饲料中β-葡聚糖的抗营养作用提供了一种有前景的生物学方法。