系统表达HB-EGF转基因小鼠的建立

The Establishment of HB-EGF Systemic Expression in Transgenic Mice

目的建立系统表达肝素结合性表皮生长因子(HB-EGF)的转基因动物模型,利用转基因动物模型研究HB-EGF与组织纤维化的关系。方法RT-PCR法克隆小鼠HB-EGF基因,将其插入Chickenβ-actin启动子下游,构建Chickenβ-actin-HB-EGF表达载体,利用显微注射的技术把表达载体注射到受精卵的雄原核中,建立HB-EGF转基因小鼠。利用特异引物PCR的方法鉴定转基因的基因型,采用Western Blot方法鉴定HB-EGF基因在全身组织的表达。分别取HB-EGF转基因鼠与同窝阴性小鼠的肝、肾、肺及膀胱组织进行Massion染色。结果建立了系统表达HB-EGF转基因小鼠,Western Blot发现其HB-EGF在肝、肺、肾及膀胱的表达与同窝阴性对照小鼠相比明显增加。Massion染色结果表明转基因鼠肝、肺、肾及膀胱组织纤维化程度明显高于同窝阴性对照小鼠。结论成功建立了系统表达HB-EGF转基因小鼠,HB-EGF的过度表可显著加重组织纤维化程度。

Objective To construct heparin-binding epidermal growth factorlike growth factor（（HB-EGF）） transgenic mice in order to investigate the role of HB-EGF in the fibrosis of tissues.Method／ HB-EGF gene was cloned form mouse with RT-PCR.The cloned HB-EGF gene was then inserted downstream of Chicken β-actin promoter to construct the vector which could express HB-EGF gene in system.The transgenic mice were produced by microinjection method and the genotype was detected by PCR with specific primers.The expression profiles of HB-EGF in body tissues were illustrated by Western Blot.Massion staining of liver,lung,kidney and bladder from transgenic mice and nontransgenic control mice was carried out.Results／ Two lines of HB-EGF transgenic mice were established.The expression levels of the HB-EGF gene in liver,lung,kidney and bladder from transgenic mice were obviously higher than those form control mice.The extents of fibrosis in the liver,lung,kidney and bladder of transgenic mice were significantly enhanced by the expression of the transgene.Conclusion／ Two lines of system-expressing HB-EGF transgenic mice were established successfully.The over-expression of HB-EGF can significantly aggravate tissues fibrosis degree.