小鼠腭裂相关基因的克隆及基因表达的意义

Cloning of the cleft palate related gene in mouse and the significance of the gene expression

目的:筛选并克隆正常组小鼠腭突与腭裂组小鼠腭突组织中差异表达的基因,探讨它们在腭裂发生过程中表达的意义。方法:建立C57BL/6N小鼠腭裂模型,分别提取胚胎12d的正常组及维甲酸处理组小鼠腭突组织中的mRNA,利用PCR的改良消减杂交技术,筛选并克隆小鼠腭突正常发育时期与腭裂形成过程中差异表达的基因及测序。结果:共获得差异表达的基因40个,2个为新基因。结论:克隆的新基因及fau基因可能对腭突间充质细胞的生长、分化具有重要的调控作用。

AIM:Cleft palate is a kind of polygenic inheritance disease. Its inheritance character base on polygene.Some of related gene to cleft palate have been found such as TGF α,TGF βand MSX.In order to explore the mechanics of the developing cleft palate,the author screen and clone the differently expression gene in palatal process between normal group and retinoic acid treated group mouse and explore the significance of the gene expression.METHODS:On gestation day 10, C57BL/6N strain mice were divided randomly into experimental and control group.On gestation day 12, mice were killed and embryo mice obtained.The mRNAs in embryo mice palatal process were isolated respectively from control and experimental group.Cloning the differently expression gene by an improved technique based on PCR subtractive hybridization and excepted false positive clone through reverse dot blot. After that their sequence were determine.RESULTS:47 differential genes were obtained. Among them there were 7 false positive clones. So 40 of differential expression gene related to mouse cleft palate were succeed clone. 26 of them were ribosomal protein gene, 2 were novel gene and 9 of them were Fau gene (Finkel Biskis Reilly murine sarcoma virus associated ubiquitously expression gene).CONCLUSION: It suggests that the new gene and fau gene were significant on the proliferation and differentiation of the mesenchymal cells in palatal process. To analysis the structure and function of the differential expression gene can make clear the base of molecular biology during developing clef1t palate.