Improved Retroviral Vectors for Gene Transfer and Expression
Improved Retroviral Vectors for Gene Transfer and Expression
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DOI：
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发表时间：
1989-10
期刊：
影响因子：
2.7
通讯作者：
Miller Ad;Rosman Gj
中科院分区：
文献类型：
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作者：
Miller Ad;Rosman Gj
我们描述了一组鼠逆转录病毒为基础的载体，包括独特的克隆位点插入cDNA，使cDNA可以驱动的逆转录病毒长末端重复序列，立即早期启动子的人巨细胞病毒，或猿猴病毒40早期启动子。该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。构建这些载体以防止剩余病毒序列合成病毒蛋白，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的同源重叠以防止辅助病毒产生。描述了用于产生高滴度病毒的方法。
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.