重组腺相关病毒-阳离子脂质体载体的构建及评价

目的探讨重组腺病毒相关病毒（rAAV）-阳离子脂质体（LyoVec）复合载体的构建方法及其转染率评价。方法构建rAAv～GFP，rAAv—LyoVec复合载体及rAAV—GFP—LyoVec。倒置荧光显微镜观察，测定rAAV—GFP，LyoVec—GFP及rAAV—GFP-LyroVec对定量C6细胞的转染率，筛选出高转染率载体。结果rAAV—GFP组24、48、72和96h转染率分别为16％、23％、21％和9％；LyoVec-GFP组为28．5％、33％、32％和11％；rAAV—GFP—Lyovec组为49％、59％、64％和20％。rAAV-GFP—LyoVec纽分别在24、48、72和96h的转染率显著高于rAAV—GFP组（P〈0．01）及LyoVec—GFP组（P〈0．01）。结论新构建rAAV—LyoVec复合载体基因的转染效率远远高于rAAV或LyoVec独立转染。

Construction and evaluation of recombinant adeno-associated virus-LyoVec vector

[Objective] To study the construction and evaluation of recombinant adeno-associated virus-LyoVec vector.[Methods] The rAAV-GFP,rAAV-LyoVec and rAAV-GFP-LyoVec were Constructed,the transfection efficiency in rAAV-GFP,LyoVec-GFP and rAAV-GFP-LyoVec to quantitative rat C6 cells were observed and measured by fluorescence microscope,and the transfection efficiency in the three vectors were confirmed and compared.The high transfection efficiency vector was be choosed.[Result] The transfection efficiency in rAAV-GFP was 16%,21%,23%,9% in 24 h,48 h,72 h,96 h;LyoVec-GFP was 28.5%,33%,32%,11%;rAAV-GFP-LyoVec was 49%,59%,64%,20%.The transfection efficiency in rAAV-GFP-LyoVec was much higher than that in rAAV-GFP(P <0.01)and LyoVec-GFP(P <0.01)respectively in 24 h,48 h,72 h,96 h.[Conclusion] The transfection efficiency of rAAV-LyoVec was much higher than the self-transfection of rAAV or LyoVec.