用于基因转移和表达的改进型逆转录病毒载体。
Improved retroviral vectors for gene transfer and expression.
作者信息
Miller A D, Rosman G J
机构信息
Fred Hutchinson Cancer Research Center, Seattle, WA 98104.
出版信息
Biotechniques. 1989 Oct;7(9):980-2, 984-6, 989-90.
Abstract
We describe a set of murine retrovirus-based vectors that include unique cloning sites for insertion of cDNAs such that the cDNA can be driven by either the retroviral long terminal repeat, the immediate early promoter of human cytomegalovirus, or the simian virus 40 early promoter. The vectors carry the neomycin phosphotransferase gene expressed from an alternate promoter as a selectable marker. These vectors have been constructed to prevent viral protein synthesis from the remaining viral sequences, to yield high-titer virus stocks after introduction into retrovirus packaging cells, and to eliminate homologous overlap with viral DNAs present in retrovirus packaging cells in order to prevent helper virus production. Methods for generating high-titer virus are described.
摘要
我们描述了一组基于鼠逆转录病毒的载体，这些载体包含用于插入cDNA的独特克隆位点，使得cDNA可以由逆转录病毒长末端重复序列、人巨细胞病毒的立即早期启动子或猿猴病毒40早期启动子驱动。这些载体携带从替代启动子表达的新霉素磷酸转移酶基因作为选择标记。构建这些载体是为了防止从剩余的病毒序列进行病毒蛋白合成，在引入逆转录病毒包装细胞后产生高滴度病毒原液，并消除与逆转录病毒包装细胞中存在的病毒DNA的同源重叠，以防止产生辅助病毒。本文描述了产生高滴度病毒的方法。
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