Many biological processes result in either cell death or cessation of cell growth. However, plasmid- and retrovirus-based mammalian expression vectors in which it has been possible to construct representative cDNA libraries cannot be readily recovered from cells that are not actively dividing. This has limited the efficiency of selection of recombinant genes that mediate either lytic events or growth arrest. Examples include genes that encode the target antigens of cytotoxic T cells, genes that promote stem-cell differentiation and pro-apoptotic genes. We have successfully constructed representative cDNA libraries in a poxvirus-based vector that can be recovered from cells that have undergone lethality-based selection. This strategy has been applied to selection of a gene that encodes a cytotoxic T-cell target antigen common to several independently derived tumors.

译文

许多生物过程导致细胞死亡或细胞生长停止。但是,基于质粒和逆转录病毒的哺乳动物表达载体 (其中可以构建代表性cDNA文库) 不能很容易地从不活跃分裂的细胞中回收。这限制了介导裂解事件或生长停滞的重组基因的选择效率。例子包括编码细胞毒性T细胞靶抗原的基因,促进干细胞分化的基因和促凋亡基因。我们已经在基于痘病毒的载体中成功构建了代表性的cDNA文库,可以从经过基于致死性选择的细胞中回收。此策略已用于选择编码几种独立来源的肿瘤共有的细胞毒性T细胞靶抗原的基因。

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