一种分离重组杆状病毒的新方法。

Patel G, Nasmyth K, Jones N

Gene Regulation Laboratory, Imperial Cancer Research Fund, Lincoln's Inn Fields, London, UK.

Nucleic Acids Res. 1992 Jan 11;20(1):97-104. doi: 10.1093/nar/20.1.97.

An improved method for the isolation of baculovirus recombinants is described. The method involves the replication and maintenance of the baculovirus genome in the yeast Saccharomyces cerevisiae which was accomplished by the isolation of a baculovirus recombinant containing yeast ARS and CEN sequences ensuring stable replication in yeast and a URA3 selectable marker. The viral DNA maintained its ability to replicate in insect cells. An efficient and rapid selection system was set up, to isolate viral recombinants in yeast; DNA from selected yeast colonies was transfected into insect cells to obtain recombinant virus. We demonstrate the utility of this system by isolating recombinant viruses that express two different members of the CREB/ATF family of transcription factors.

本文描述了一种改进的杆状病毒重组体分离方法。该方法涉及杆状病毒基因组在酿酒酵母中的复制和维持，这是通过分离一种含有酵母自主复制序列（ARS）和着丝粒（CEN）序列的杆状病毒重组体来实现的，这些序列确保了在酵母中的稳定复制以及一个URA3选择标记。病毒DNA保持了其在昆虫细胞中复制的能力。建立了一个高效快速的选择系统，用于在酵母中分离病毒重组体；将从选定酵母菌落中提取的DNA转染到昆虫细胞中以获得重组病毒。我们通过分离表达CREB/ATF转录因子家族两个不同成员的重组病毒，证明了该系统的实用性。