多药耐药基因-腺病毒重组表达载体的构建及鉴定

Construction and identification of multi-drug resistance gene-adenovirus recombinant expression vector

目的构建多药耐药(multidrug resistance,MDR)基因-腺病毒重组表达载体。方法将MDR1基因插入到腺病毒载体,用磷酸钙沉淀法将此重组载体转染到293细胞,制备重组病毒悬液并感染靶细胞进行鉴定。结果构建的MDR1-腺病毒载体在293细胞中大量产生,病毒滴度达109PFU/ml,此病毒感染靶细胞后,在靶细胞中有MDR1基因的表达。结论成功构建了MDR1-腺病毒重组表达载体。

Objective Construction of multi-drug resistance gene(MDR)-adenoviral recombinant expression vector.Methods MDR1 gene will be inserted into the adenovirus vector,by calcium phosphate precipitation method the recombinant plasmid transfect into 293 cells.Recombinant virus suspension is made and infeced target cells and to indentify.Results Constructed MDR1-adenovirus vector produces a large number in 293 cells,the virus titer is up to 109 PFU/ml.After the recombinant virus infected target cells in the target cells have the expression of MDR1 gene.Conclusion MDR1-recombinant adenorirus vector has been successfully constructcd.