Caputo A, Barbanti-Brodano G, Reschiglian P, Gianni M, Mottes M, Miranda P, Milanesi G, Knowles B B, Ricciardi R P
Institute of Microbiology, University of Ferrara, Italy.
J Gen Virol. 1988 Feb;69 ( Pt 2):459-66. doi: 10.1099/0022-1317-69-2-459.
The construction of the first stable human cell lines that express and secrete authentic hepatitis B virus surface antigen (HBsAg), using a BK virus (BKV) episomal plasmid vector, is described. The amount of HBsAg produced by BKV vectors (up to 600 ng/10(7) cells) was comparable to other eukaryotic vector systems. The level of HBsAg expression remained the same regardless of the orientation of the HBsAg gene, substitution of the HBsAg gene promoter with the mouse metallothionein I gene promoter or the tissue origin of the human cell lines used to establish stable cellular transformants. Northern blot analysis also indicated synthesis of normal HBsAg transcripts. Surprisingly, however, the vectors were maintained at far lower than expected copy number (one to five copies/cell). Reasons for this are discussed.
本文描述了利用BK病毒(BKV)游离体质粒载体构建首个稳定表达并分泌真性乙型肝炎病毒表面抗原(HBsAg)的人细胞系。BKV载体产生的HBsAg量(高达600 ng/10⁷个细胞)与其他真核载体系统相当。无论HBsAg基因的方向如何、用小鼠金属硫蛋白I基因启动子替代HBsAg基因启动子,还是用于建立稳定细胞转化体的人细胞系的组织来源如何,HBsAg的表达水平均保持不变。Northern印迹分析也表明合成了正常的HBsAg转录本。然而,令人惊讶的是,载体维持在远低于预期的拷贝数(1至5个拷贝/细胞)。文中讨论了其原因。