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为了抑制Tat蛋白的反式激活作用,在细胞内大量表达外源TARRNA使其与Tat蛋白结合,从而竞争性抑制其与HIV-1LTR的TARRNA元件结合.构建了以HIV-1LTRYL158(-158~+180)为启动子,分别含有4,8和15个拷贝的TAR-CoreRNA诱饵(decoy)表达质粒;以荧光酶基因为报告基因,检测了瞬时共转染体系中含不同拷贝数的TAR-CoreDNA转录产物对Tat蛋白反式激活作用的影响.结果证明,TAR-CoreRNA诱饵对Tat蛋白活性具有很强的抑制作用,其抑制程度与TAR-CoreDNA串联体的拷贝数有关.
In order to inhibit the transactivation of Tat protein, exogenous TARRNA is expressed in the cell in large quantities to bind to the Tat protein, thereby competitively inhibiting its binding to the TARRNA element of HIV-1 LTR. A decoy expression plasmid containing 4, 8 and 15 copies of TAR-CoreRNA was constructed with HIV-1LTRYL158 (-158 ~ + 180) as a promoter. The luciferase gene was used as a reporter gene to detect transient co-transfection Effects of different copies of TAR-CoreDNA transcripts on the transactivation of Tat protein in the system. The results demonstrated that the TAR-CoreRNA decoy has a strong inhibitory effect on the Tat protein activity, and the degree of inhibition is related to the copy number of the TAR-Core DNA concatemer.