反义RNA对细胞系CC81中牛白血病病毒复制的抑制作用
Inhibition of bovine leukaemia virus replication by the antisense RNA in cell line CC81.
作者信息
Borisenko A, Miroschnickhenko O, Tikchonenko T I
机构信息
Institute of Agricultural Biotechnology, Moscow, Russia.
出版信息
Virus Res. 1992 Apr;23(1-2):89-97. doi: 10.1016/0168-1702(92)90069-l.
A model system has been developed for quantitative evaluation of bovine leukaemia virus (BLV) replication in a permanent cell line CC81. Transfection of the BLV DNA into these cells evoked typical signs of retroviral infection: formation of syncytia, manifestation of reverse transcriptase activity and appearance of characteristic budding retroviral particles. To inhibit BLV replication, a recombinant plasmid pAGR with an antisense RNA gene targeted at the R-U5 region (147th-342th nt) of the viral genome has been engineered. Cotransfection of CC81 cells with infectious BLV DNA and pAGR led to effective inhibition of BLV replication by the antisense RNA, evidenced by a drop in the number of syncytia and reverse transcriptase activity. Maximal inhibition of BLV replication (95-97%) was observed at a weight ratio of input viral and plasmid DNAs equal to 1:10.
已开发出一种模型系统，用于定量评估牛白血病病毒（BLV）在永久细胞系CC81中的复制情况。将BLV DNA转染到这些细胞中引发了逆转录病毒感染的典型迹象：形成多核巨细胞、逆转录酶活性的表现以及特征性出芽逆转录病毒颗粒的出现。为了抑制BLV复制，构建了一种重组质粒pAGR，其带有靶向病毒基因组R-U5区域（第147至342个核苷酸）的反义RNA基因。用感染性BLV DNA和pAGR共转染CC81细胞导致反义RNA对BLV复制的有效抑制，这通过多核巨细胞数量和逆转录酶活性的下降得以证明。当输入的病毒DNA与质粒DNA的重量比等于1:10时，观察到BLV复制的最大抑制（95-97%）。
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