腺相关病毒与人多药耐药基因重组载体的构建及在NIH3T3细胞中的表达

腺相关病毒 (adeno- associated virus,AAV)属细小病毒科 ,是一种最小的动物病毒 .具有其他病毒载体所没有的优点 ,在基因治疗中日益受到瞩目 .以 AAV的一种多克隆载体为基础 ,构建了携带 MDR1基因的重组腺相关病毒载体 (r AAV- MDR1 ) ,经 2 93细胞包装成重组病毒 .将重组质粒、重组病毒分别转染和感染 NIH3T3细胞 ,用 PCR和 MTT法检测了人 MDR1基因的转导及表达 .为 MDR1基因用于临床和腺相关病毒载体在基因治疗中的应用提供了依据

Construction of Recombinant AAV-MDR1 and Expression in NIH3T3 Cells

Transfer of the human multidrug resistance 1 (MDR1) gene to hematopoietic stem cells offers an approach to overcome the myelosuppression caused by a number of antineoplastic drugs.Adeno associated virus is a human parvovirus with many interesting features in its life cycle that can be exploited in human gene therapy.So it was chosen as a vector in the hope of developing a safe and efficient viral vector for human gene therapy.This study was designed to construct the recombinant AAV MDR1 vector and to package the AAV MDR1 vector into viruses.The polymerase chain reaction (PCR) based assay was used to determine the existence of MDR1 and MTT method to determine the expression of MDR1 in NIH3T3 cells.The results show that AAV can mediate MDR1 gene into NIH3T3 cells.