Murovska M F, Chernobayeva L G, Miroshnichenko O I, Tomsons V P, Konicheva V V, Ivanova S V, Tikhonenko T I
Institute of Microbiology, Latvian Academy of Sciences, Riga.
Vet Microbiol. 1992 Nov;33(1-4):361-6. doi: 10.1016/0378-1135(92)90063-y.
A possible approach to control of bovine lymphoproliferative disease caused by bovine leukaemia virus (BLV) may be the development of an "antiviral information immunity" based on the effect of anti-sense RNA (asRNA). A numbers of constructs were obtained, under control of various promotors (herpesvirus thymidine kinase, T-antigen SV40 promoter), carrying as DNA against gene X, the expression product of which is a transactivator of viral transcription from the BLV LTR promotor. As a model system for the analysis of antiviral activity of constructs developed, cloned continuous cell lines of BLV-producing FLK cells were used. The level of BLV expression in cells transfected with the constructs was determined by various parameters. Differences were detected in different clones obtained from non-transfected cells, as well as variation between transfected clones, as measured by reverse transcriptase, competitive radio-immunoassay for BLV p24, the viral particle count on agar membrane, and the tumorigenicity for nude mice. The differences in inhibition of expression of BLV genes and their products may be explained in terms of the site of integration of asDNA and the number of integrated copies.
控制由牛白血病病毒(BLV)引起的牛淋巴增生性疾病的一种可能方法,或许是基于反义RNA(asRNA)的作用来开发一种“抗病毒信息免疫”。在各种启动子(疱疹病毒胸苷激酶、SV40 T抗原启动子)的控制下,获得了许多构建体,它们携带针对基因X的DNA,该基因的表达产物是来自BLV LTR启动子的病毒转录反式激活因子。作为分析所开发构建体抗病毒活性的模型系统,使用了产生BLV的FLK细胞的克隆连续细胞系。通过各种参数确定构建体转染细胞中BLV的表达水平。通过逆转录酶、BLV p24竞争性放射免疫测定、琼脂膜上的病毒颗粒计数以及裸鼠的致瘤性来衡量,从未转染细胞获得的不同克隆以及转染克隆之间均检测到差异。BLV基因及其产物表达抑制的差异或许可以根据反义DNA的整合位点和整合拷贝数来解释。