转manA基因小鼠F_1代表达效果的检测分析  

Analysis of transgene expression and effect on F_1 generation of manA transgenic mice

作　　者：张茂[1] 李紫聪[2] 许惠[2] 张冠冠[2] 刘德武[2] 吴珍芳[2] 

机构地区：[1]龙岩学院生命科学学院/预防兽医学与生物技术福建省高等学校重点实验室,福建龙岩364012 [2]华南农业大学动物科学学院,广东广州510642

出　　处：《广东农业科学》2014年第9期147-150,154,共5页Guangdong Agricultural Sciences

基　　金：广东省科技计划项目(2011A020102003);广东省农业攻关项目(2011A020201009)

摘　　要：将携带β-甘露聚糖酶基因manA的转基因FVB原代小鼠与野生型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β-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β-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。The β-mannanase manA transgene FVB mice were mated with the wild-type FVB mice to gain the F1 generation 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 RTPCR detection and quantitative real-time PCR. The result showed that manA was specific expressed in parotid and sublingual glands, and was highest expressed in the sublingual gland of the transgenic mice 302 line, certificating that manA was successfully specific expressed in the parotid gland and sublingual gland of transgenic mice. Also, the saliva and dejecta of the F1 mice were collected to detect the activity of mannanase and digestibility of nutrients in feed. The result showed that the 302 line mice had the highest enzymatic activity, digestibility of crude fat in 302 line mice was significantly different with wild-type mice, while no significant difference in digestibility of crude protein and crude fiber between them. There were no significant differentce in crude protein, crude fate and crude fiber between 304 line mice and wild-type mice.

关 键 词：转基因 manA基因 检测 分析 

分 类 号：Q786[生物学—分子生物学]