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- W2380102142 abstract "In order to facilitate gene therapy research using recombinant adeno associated virus(rAAV) vector as gene transfer vehicle, we constructed a series of rAAV vectors including pWAV 1, pWAV 2 and pSNAV for inserting foreign genes, each of which is composed of the two ITRs of AAV 2 and in between are the human cytomegalovirus (CMV) immmediate early enhancer and promoter, multiple cloning sites(MCS) and polyadenylation signal. A rAAV vector carrying a gene expression cassete can be conveniently obtained by inserting the target gene into the MCS in a correct direction. The MCS of pWAV 1 includes NheⅠ,XhoⅠ,PstⅠand BamHⅠ, where up to 3.3kb foreign genes can be inserted for packaging of rAAV particles. Both pWAV 2 and pSNAV have the same capacity of inserting 3.6kb foreign genes, with MCS of KpnⅠ, EcoRⅠ, SalⅠ for the former and KpnⅠ, EcoRⅠ, SalⅠ and BglⅡ for the latter. We also constructed rAAV vectors pAV lacZ and pSNAV lacZ, both harboring β galactosidase gene. After transfecting pAV lacZ and pSNAV lacZ into BHK 21 cells respectively, a recombinant herpes simplex virus(rHSV) having the whole helper functions for rAAV packaging that we reported recently was used to infect the cells. Two strains of rAAV carrying β galactosidase gene were obtained with a titer of 5×10 4 TU/ml respectively, which not only confirmed the feasibility of this method but also indicated the possibility to produce rAAV in large scale using “one helper virus one vector cell line” strategy." @default.
- W2380102142 created "2016-06-24" @default.
- W2380102142 creator A5075548449 @default.
- W2380102142 date "2000-01-01" @default.
- W2380102142 modified "2023-09-28" @default.
- W2380102142 title "Construction of a Series of Adeno associated Virus Vectors and Their Expression of β-galactosidase Gene" @default.
- W2380102142 hasPublicationYear "2000" @default.
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