摘要
目的 探讨慢病毒载体介导的截短肌营养不良蛋白cDNA表达的可行性。方法 将PCR克隆所构建的三个截短的肌营养不良蛋白cDNA插入到慢病毒载体,再将此载体转染至293Ad^3+细胞中包装为重组病毒。用重组病毒感染培养的鼠成肌细胞,通过蛋白免疫印迹检测鼠成肌细胞中截短肌营养不良蛋白cDNA的表达。结果 用PCR克隆所构建的三个截短肌营养不良蛋白cDNA通过慢病毒载体导入鼠成肌细胞后均可表达出特异产物。结论 慢病毒载体可介导截短肌营养不良蛋白cDNA的表达,截短肌营养不良蛋白cDNA和慢病毒载体分别有望作为基因治疗Duchenne肌营养不良的目的基因和载体。
Objective To explore the expression feasibility of truncated dystrophin cDNAs mediated by lentiviral vector. Methods Three truncated dystrophin cDNAs were constructed by PCR cloning and inserted into lentiviral vector. The recombinant vectors were transiently transfected into 293Ad^5+ cells. The recombinant lentiviruses infected the cultured myoblasts from rats. Expressions of truncated dystrophin cDNAs were detected by Western blot analysis. Results Mediated by lentiviral vector, three cDNAs constructed by PCR cloning expressed relative truncated dystrophins in the transfected myoblasts. Conclusion Truncated dystrophin cDNAs can be expressed successfully under the mediation of lentiviral vector, which offers the possibility of utilizing truncated dystrophin cDNAs and lentiviral vector for gene therapy of Duchenne muscular dystrophy.
出处
《中华神经医学杂志》
CAS
CSCD
2007年第2期148-151,共4页
Chinese Journal of Neuromedicine