RU5区对BFV3026基因表达的调控

Regulation of RU5 Region to Gene Expression of BFV 3026

以牛泡沫病毒(Bovine foamy virus, BFV)中国株BFV3026原病毒DNA为材料,构建R区系列缺失质粒, 通过对其转染细胞中RT水平及对缺失质粒与luc报告质粒共转染细胞中萤火虫荧光素酶活性的测定,确立U5区对于BFV3026两类启动子LTR和IP均具有负调控作用;同时将带有不同R区的BFV3026结构基因片段克隆于异源启动子CMV之下,通过对其转染细胞293T中RT酶活性的测定,确立R区对于病毒结构基因pol的表达具有一定的调节作用,并将其功能区域初步界定在R区5′端100bp内。

In order to elucidate the regulation mechanism of RU5 region to BFV gene expression, BFV3026 provirus DNA was used to construct the plasmids containing different deletion of R region, which were cotransfected with luc report gene locatied behind the IP promoter to BL12 cells, and luciferase activities was assayed, confirming that U5 region could repress the initiation function of LTR as well as IP. The BFV structure genes with different deletion of R region were cloned closely behind to heterogenous CMV promoter, then transfected to 293T cells, RT activity was performed, testifying the R region was required for BFV pol gene expression, and also the function domain was identified within the 100n.t. sequence at the 5′end.