ICR鼠皮肤Sonic Hedgehog(Shh)基因的克隆及表达  

Clone and expression of Sonic Hedgehog (Shh) gene from embryonic mice skin

作　　者：朱晓芳[1,2] 王琴 成蓓 高慧 原志伟[4] 朱国强[4] 范卫新[1] 

机构地区：[1]南京医科大学第一临床医学院皮肤科,210029 [2]扬州大学临床医学院皮肤科,扬州225001 [3]扬州大学临床医学院 [4]扬州大学兽医学院

出　　处：《中国麻风皮肤病杂志》2009年第9期665-667,共3页China Journal of Leprosy and Skin Diseases

基　　金：国家自然科学基金(30771947);江苏省自然基金(BK2007248);扬州市科技攻关项目基金(YZ2008041)

摘　　要：目的:检测ICR胎鼠皮肤组织克隆Sonic Hedgehog(Shh)基因在胎鼠皮肤的表达。方法:分离12.5～16.5 d胎龄的胎鼠背部皮肤,用Trizol-酚-氯仿抽提法提取总RNA,应用RT-PCR技术扩增胎鼠皮肤Shh全部编码基因,重组于pMD19-T载体进行克隆测序分析。应用合成地高辛标记的反义Shh-mRNA探针,整体原位杂交技术检测Shh基因在ICR鼠皮肤上的表达。结果:从15.5 d胎鼠背部皮肤中成功扩增Shh基因,Blast分析与GeneBank公布的Shh基因编码区一致。同时15.5 d以后胎鼠背部皮肤检测到Shh基因的表达。结论:Shh基因可能参与了皮肤毛囊的形成。Objective： To measure the clone and expression of Sonic Hedgehog （Shh） gene from embryonic ICR mice skin. Methods： The total RNAs of E12.5 - 16.5 day mice skin were obtained by Trizol - phenol - chloroform reagents. Reverse transcription PCR was used to amplify the all encoded sequence of Shh gene. The amplified fragment was cloned into pMD19 - T vector and sequenced. The anti - sense Shh mRNA probe was labeled with synthesized digoxin, and the Shh gene expression on skin detected using whole- body in situ hybridization. Results： The cDNA of Shh was obtained from embryonic mice skin. Blast analysis showed that the sequence was identical to that of GeneBank. Shh mRNA expression was detected on the 15.5 day mice back skin. Conclusion： Shh may be involved in the morphogenesis of embryonic mice hair follicle.

关 键 词：Sonic Hedgehog(Shh) 克隆 基因表达 ICR鼠 

分 类 号：R751[医药卫生—皮肤病学与性病学]