| 张茂2，李紫聪1，许惠1，张冠冠1，刘德武1，吴珍芳1.转manA基因小鼠F1代表达效果的检测分析[J].广东农业科学,2014,41(9):147- |
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转manA基因小鼠F1代表达效果的检测分析 |
| Analysis of transgene expression and effect on F1generation of manA transgenic mice |
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| DOI： |
| 中文关键词: 转基因 manA 基因 检测 分析 |
| 英文关键词: transgene manA gene detection analysis |
| 基金项目:广东省科技计划项目（2011A020102003）；广东省
农业攻关项目（2011A020201009 |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| The 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. |
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