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牛乳头瘤病毒等离子体维护序列的特征

M Lusky, M R Botchan

|February 1, 1984

概括

牛乳头瘤病毒 (BPV-1) 区域使细胞中基因的染色体外复制成为可能. 这些等离子体维护序列 (PMS) 在存在病毒因素时防止集成,确保稳定的核等离子体维护.

科学领域:

分子生物学分子生物学
病毒学病毒学
哺乳动物细胞培养

背景情况:

牛乳头瘤病毒 (BPV-1) 存在于哺乳动物细胞中的多副本核等离子体.
了解BPV-1的复制和维护机制对于基因传递和载体发展至关重要.

研究的目的:

确定和描述BPV-1基因组中负责染色体外复制和稳定等离子体维护的区域.
研究BPV-1基因产品在防止病毒DNA集成中的作用.

主要方法:

利用Tn5新菌素抗性基因作为复制研究的标记.
在BPV-1基因组内定位的等离子体维护序列 (PMS-1和PMS-2).
在各种细胞系中构建并测试了具有PMS元素的重组等离子体,包括那些提供BPV-1因子的转基因和那些缺乏它们的细胞系.
生成和分析BPV-1删除突变体,以评估特定病毒基因产物的作用.

主要成果:

在BPV-1基因组内,两个不同的区域 (PMS-1和PMS-2) 独立地支持标记基因的染色体外复制.
在没有选择性压力的情况下,PMS元件便于在恒定拷贝数下保持稳定,未经重新排列的等离子体维护.
转化所必需的BPV-1基因产物不需要维护等离子体.
在存在转基因病毒因子时,BPV-1 DNA 保持为非集成的核等离子体,而共同转基因标记基因集成到宿主基因组中.

结论:

BPV-1 具有特定的信号,促进了染色体外复制和稳定的核等离子体维护.
由于存在BPV-1转基因,病毒基因组不能被积极地集成到宿主细胞DNA中.
这些发现对开发基于BPV-1的载体对哺乳动物细胞中稳定基因表达有影响.

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