The Study Of The Production Of Plasmid Helper Recombinant Adeno-associated Virus 2 And The Reconstruction Of AAV2 To Fit The Gene Therapy Of Cerebral Ischemia And Astrocytoma.

Following the finish of the plan of human genome and the biginning of the study of the functional gene, more and more genes were used to treat human disease. Gene therapy became the most powerful tool to cure misfortune disease. The key technology of gene theapy was the choise of the vector to transfer gene, which needed to express highly in recipient cells. It is always difficult to choise a suitable vector for gene therapy. Although as the concept of virous vector had been presented from seventies of last century, various kinds of defective virous vector have been found and applied extensively. The nervous toxin of herpes simplex vector , the immunity of adeno-virus and the carcinogenesis of retrovirus limit them application in central nervous system. rAAV was attracted at high safety,extensive host,high efficiency of transfection and other prominent superiorities, which was the most suitable vector forgene therapy of central nervous system at present.The producion of rAAV was difficult, which was another barrier for application of rAAV. The methods to produce rAAV was complex and needed expensive equipments, which couldn't present in a common library. How to optimize a simple method to produce rAAV, which can finish in common library would promote application of rAAV in gene therapy.The common CMV promoter rAAV was not suitable for gene therapy of cerebral vascular disease and glioma. In cerebral vascular disease, rAAV expressed aimed gene for a long time, and leaded to hyperplasia of vessels and enhanced growth of tumor. In central nervous system, neuron had high affinity to rAAV than glial cells, which would lead to need an specific promoter to control toxin gene expression in glioma therapy.Therefore, this study was focus on the production of plasmid helper recombinant adeno -associated virus 2 and the reconstruction of AAV2 to fit the gene therapy of cerebral ischemia and astrocytoma.the production of plasmid helper recombinant adeno -associated virus 2: Plasmid helper rAAV system was transferred to HEK293T cells through phosphorated...