牦牛乳头瘤病毒I型BgPV-1转录图谱的构建及解析

The production performance of yaks directly affect the living standard in the Qinghai-Tibetan Plateau. The papillomatosis, which is caused by papillomavirus, is one of the important infectious disease affecting the production performance of yaks and occurred frequently during last several years. It can cause mastitis, bleeding during milking and distortion of the milk, and interfere with suckling and milking, and also cause reproduction problems and decline of fur quality. In our previous study, the first papillomavirus strain of yaks was identified and was designated as Bos grunniens papillomavirus type 1 (BgPV-1) by The International Committee on Taxonomy of Viruses. However, the transcription mechanism and other characters of this virus are not studied yet since there is no effective isolation and culture methods for papillomavirus. Therefore, we plan to construct a full transcription map for BgPV-1. Raft cultures derived from yak keratinocytes with BgPV-1 infection and transinfected by BgPV-1 infectious clone will be prepared respectively. Then the start sites and polyadenylation sites for early and late transcripts, splice junctions and other potential promoters in the early and late region, and what regions of the viral genome transcribed will be revealed by 5'/3' RACE and RNA-seq. The results will assist our understanding of BgPV-1 for molecular biological characterization.

牦牛的生产性能直接决定了藏区人民的生活水平，近年来牦牛的乳头瘤病频发，造成乳房炎、哺乳困难、繁殖障碍和毛皮质量下降等问题。本团队在青海省成功地鉴定到了世界上第一株牦牛乳头瘤病毒，被国际病毒分类委员会命名为牦牛乳头瘤病毒I型（BgPV-1）。但由于乳头瘤病毒无法进行体外分离培养的特点，使得BgPV-1的转录方式等尚未得到深入的研究。本研究围绕BgPV-1转录图谱的构建及其转录方式有何特点的科学问题，通过建立从病变部位分离的含有BgPV-1基因组的角质形成细胞的筏式培养体系，以及建立DNA感染性克隆转染后的筏式细胞培养体系，分别对病毒转录后的mRNA采用5'/3' RACE和RNA-seq的分析方法，确定BgPV-1早期和晚期基因转录的起始位点、剪接位点、潜在启动子、多腺苷酸化裂解位点、以及基因组的准确转录区域等，构建BgPV-1的转录图谱。为更好地全面解析该病毒的分子生物学特性奠定基础。