Expression and effect of recombinant adeno-associated virus vector-8 transduced green fluorescent protein gene into rat keratocytes
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R772.2

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National Natural Science Funds for Young Scholars(No.30901650); Natural Science Foundation of Guangdong Province, China(No.10451051501005773)

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    Abstract:

    ·AIM:To evaluate the expression and the effect of recombinant adeno-associated virus(rAAV)vector-8 transduced enhanced green fluorescent protein(EGFP)gene into rat keratocytes.·METHODS:rAAV8-EGFP was transfected respectively into rat keratocytes at different multiplicities of infection.The inverted fluorescent microscope was performed to assess GFP expression and the percentage of positive GFP in the cells was analyzed by flow cytometric analysis.The effects of rAAV8 and rAAV8-EGFP on cell proliferation were assessed by methyl thiazolyl tetrazolium(MTT)method.·RESULTS:The inverted fluorescent microscope was performed to assess GFP expression.The EGFP expression peak occurred at 7d after transfection and the expression level was 31.5%(MOI=5×103),42.5%(MOI=5×104),54.8%(MOI=5×105),respectively.Results of MTT assay indicated there was no inhibitory effect on keratocytes by rAAV8-EGFP and rAAV8 transduction respectively.·CONCLUSION:The rAAV8 vector can deliver effectively EGFP genes into rat keratocytes,and there is no inhibitory effect on keratocyte by rAAV8-EGFP transduction.·

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Ming-Fang Tang, Jian Yu, Lang Bai, et al. Expression and effect of recombinant adeno-associated virus vector-8 transduced green fluorescent protein gene into rat keratocytes. Guoji Yanke Zazhi( Int Eye Sci) 2011;11(5):769-771

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