中文摘要：

目的 ：建立携带突变位点R258C的平滑肌肌动蛋白α（ACTA2）基因的转基因小鼠模型。方法：利用基因重组技术,将人工合成的小鼠ACTA2基因第七外显子258位精氨酸（R）突变为半胱氨酸（C）,再克隆到质粒p Bluescript KS的多克隆位点,通过显微注射法,把线性化、纯化后含有鼠ACTA2启动子/外显子1/内含子1/外显子2~9（含R258C）/3′端非翻译区的质粒注射入C57BL/6J小鼠受精卵中,将存活的胚胎移植入同期发情的假孕受体母鼠内,获得子代小鼠。用聚合酶链反应（polymerase chain reaction,PCR）和Southern印迹杂交方法检测子代鼠尾DNA,鉴定基因型。结果：移植注射胚胎后得到32只新生仔鼠,经PCR和Southern印迹杂交检测得到3只阳性小鼠。结论：携带突变位点R258C的ACTA2基因的转基因鼠模型构建成功,为进一步探索ACTA2基因突变在胸主动脉瘤发病中的作用和机制提供了重要的前期工作基础。

英文摘要：

Objective： To construct the mouse model of ACTA2 mutation transgenic mice. Methods： Mutation of ACTA2 gene was created by site-directed mutagenesis. Recombinant DN A technique was used for the construction of ACTA2 promoter-/exon1/intron1/exon2-9（R258C） /3UTR transgene. The gene plasmids were microinjected into the fertilized oocytes, and then the alive zygote cells were transplanted into the pseudopregant mice. The genotype of transgenic descendants was identified by PCR and Southern blot. Results： Thirty-two new-born mice were obtained by introcytoplasmic sperm injection-embryo transplantation. Three of the 32 mice were demonstrated as ACTA2 mutation transgenic mice by PCR and Southern blot. Conclusions： ACTA2 R258 C mutation transgenic mouse model is successfully established.This model may provide a basis for studies of the roles and mechanisms of ACTA2 in the pathogenesis of thoracic aortic aneurysm.