The Experimental Research of rAAV2/1 Vector Mediated Transfection of Enhanced Green Fluorescent Protein on the Sacral Cord
HU Yingqiu
XU Meiling
Abstract：To recombine adeno -associated virus vector (recombinant adeno -associated virus,rAAV)carrying enhanced green fluorescent protein (enhanced green fluorescent protein,EGFP)gene into the rat sacral cord,observe the location and expression and evaluate it as the feasibility of gene therapy vector of sacral cord lesion.SD rats were injected rAAV2 /1 -EGFP in bladder muscle by more,according to different multiplicity of infection (1 ×109、1 ×10 10、4 ×10 10 ).The rats were deeply anesthetized after two weeks, four weeks,eight weeks and fixed by 4% paraformaldehyde with the left ventricle -aorta cannulation,the spinal cord was slowly stripped and put into the PBS containing 30% sucrose in the night (4 ℃)and serial sectioned within the frozen section machine at constant temperature,the thickness was 12 μm and stained with immunofluorescence (GFP antibody,FITC labeled goat anti -rabbit IgG),the EGFP fluorescence expression was observed under the microscope.Fluorescence microscopy revealed that four weeks later,the multiplicity of infection of 4 ×1010 had strong fluorescence of sacral cord neurons,showing the enhanced expression of green fluorescent protein diffusion.EGFP gene can be effectively transfected into rat sacral cord by rAAV2 /1 vector ,rAAV2 /1 vector is an ideal gene ther-apy vectors.For rAAV2 /1 vector is transfected as the treatment of sacral nerve -born diseases and provide a theoretical basis.
Keywords：Recombinant adeno -associated virusEnhanced green fluorescent proteinsacral cordTransfectionGene therapy
Publication Date：2013-01-01
Online Publishing Date：2026-09-11(First online date of this platform, not the publication date of the document)
Pages：4( 239-242 )