转manA基因小鼠F1代表达效果的检测分析

将携带茁-甘露聚糖酶基因manA 的转基因FVB 原代小鼠与“生型FVB 小鼠进行繁殖得到F1代，通过PCR 和Southern blot 鉴定出F1代转基因小鼠。提取转基因小鼠腮腺、舌下腺、下颌下腺、心脏、肝脏等组织的总RNA，通过RT-PCR 检测出manA 在转基因小鼠的腮腺和舌下腺组织特异性表达。进行定量PCR 验证出manA 基因在302号家系小鼠的舌下腺中表达量最高，证明茁-甘露聚糖酶基因manA 在转基因小鼠的腮腺和舌下腺特异性表达成功。对转基因小鼠唾液进行收集测定茁-甘露聚糖酶活性，在302 号家系中检测到酶活性较高。通过代谢试验测定饲料中粗蛋白、粗纤维和粗脂肪的消化率，结果显示，与非转基因小鼠相比，302 号家系转基因小鼠对饲料中粗蛋白、粗纤维的表观消化率无明显差异，而粗脂肪的消化率得到显著提高；304 号家系转基因小鼠对饲料中粗蛋白、粗脂肪、粗纤维消化率方面均无明显差异。

Analysis of transgene expression and effect on F1generation of manA transgenic mice

The mannanase manA transgene FVB mice were mated with the wild-type FVB mice to gain the F1generation transgene mice that identified by PCR and Southern blot. Total RNA of parotid gland, sublingual gland,submandibular gland, heart, liver and other tissues of the F1 generation transgene mice were extracted to perform the RTPCRdetection and quantitative real -time PCR. The result showed that manA was specific expressed in parotid andsublingual glands, and was highest expressed in the sublingual gland of the transgenic mice 302 line, certificating thatmanA was successfully specific expressed in the parotid gland and sublingual gland of transgenic mice. Also, the saliva anddejecta of the F1 mice were collected to detect the activity of mannanase and digestibility of nutrients in feed. The resultshowed that the 302 line mice had the highest enzymatic activity, digestibility of crude fat in 302 line mice wassignificantly different with wild-type mice, while no significant difference in digestibility of crude protein and crude fiberbetween them. There were no significant differentce in crude protein, crude fate and crude fiber between 304 line mice andwild-type mice.