| 王刚 方诚 李先贵 王飞 赵占伟 杜建军.GDDR 转基因小鼠的基因表达鉴定及功能初步分析[J].现代生物医学进展英文版,2015,15(10):1836-1839. |
| GDDR 转基因小鼠的基因表达鉴定及功能初步分析 |
| The Gene Expression Identification and Elementary Functional Analysisof GDDR Transgenic Mice |
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| DOI： |
| 中文关键词: GDDR 转基因小鼠 鉴定 功能分析 |
| 英文关键词: GDDR Transgenic mice Identification Functional analysis |
| 基金项目:国家自然科学基金项目（81270440） |
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| 中文摘要: |
| 目的：对构建的胃癌相关下调基因GDDR（Gastric Dramatic Down-related）转基因小鼠进行鉴定及功能初步检测，为深一步
研究GDDR的生物学功能提供可靠的动物模型。方法：对GDDR转基因和野生对照（未经过基因修饰）小鼠胃组织使用实时定量
PCR 检测其基因转录情况，western blot和免疫组化检测其组织蛋白表达水平及表达部位。通过胃组织进行糖原染色和胃粘膜表
面主要糖蛋白MUC5AC 行免疫组化染色对小鼠表型进行初步分析。结果：实时定量PCR、western blot和免疫组化结果均显示在
GDDR 转基因小鼠中胃粘膜GDDR的表达水平明显高于野生组。糖原染色结果显示GDDR 转基因小鼠的粘液分泌及粘液厚度
高于正常对照组，糖蛋白MUC5AC的表达也显著强于野生组。结论：经过鉴定GDDR转基因小鼠的外源性基因GDDR得到成功
表达。通过对GDDR转基因小鼠表型的初步分析，提示GDDR 可能参与胃粘膜粘液层的组成和增加粘液分泌从而发挥着保护胃
粘膜的作用。 |
| 英文摘要: |
| Objective:To provide a reliable animal model for the study of deep step GDDR （Gastric Dramatic Down-related）
biological functions, the identification and primary functional study of GDDR transgenic mice were performed.Methods:Quantitative
real-time PCR was used to detect transcriptional level of gene GDDR in GDDR transgenic mice and the wildtype (gene without any
modifications). Western blot and immunohistochemistry were used to identify protein expression level and expression site. Periodic
Acid-Schiff stain（PAS）for gastric mucus secretion and immunohistochemical staining for major glycoprotein MUC5AC in the surface
of gastric mucosa were used to make the primary functional analysis of GDDR transgenic mice.Results:Quantitative real-time PCR,
western blot and immunohistochemistry showed that the level of gene GDDR expression in gastric mucosa was significantly higher in
GDDR transgenic mice than in the wildtype. Periodic Acid-Schiff stain showed that mucus secretion and the thickness of mucus in
GDDR transgenic mice was significantly higher than the normal control group. The expression of glycoprotein MUC5AC in the gastric
mucosa of GDDR transgenic mice was also significantly stronger than the wildtype.Conclusion:Exogenous gene GDDR in GDDR
transgenic mice was successfully expressed after identification .By making preliminary analysis of gene GDDR function in transgenic
mice, it is considered that GDDR may play an important role in gastric mucosal protection through participating in the composition of
gastric mucus layer and contributing to mucus secretion. |
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