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High-titer packaging cells producing recombinant retroviruses resistant to human serum.

机译:高滴度包装细胞产生抗人血清的重组逆转录病毒。

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摘要

Novel retroviral protein expression constructs were designed to retain minimal retroviral sequences and to express dominant selectable markers by reinitiation of translation after expression of the viral genes. HT1080 cells were selected as producer cells for their ability to release high-titer viruses that are resistant to inactivation by human serum. Two HT1080-based packaging cell lines which produce Moloney murine leukemia virus cores with envelope glycoproteins of either amphotropic murine leukemia virus (FLYA13 line) or cat endogenous virus RD114 (FLYRD18 line) are described. Direct comparison with previous retroviral packaging systems indicated that 100-fold-higher titers of helper-free recombinant viruses were released by the FLYA13 and FLYRD18 lines.
机译:设计新颖的逆转录病毒蛋白表达构建体,以保留最小的逆转录病毒序列,并在表达病毒基因后通过重新启动翻译来表达显性选择标记。选择HT1080细胞作为生产细胞,因为它们具有释放高滴度病毒的能力,这些病毒对人血清的灭活具有抵抗力。描述了两种基于HT1080的包装细胞系,它们产生莫洛尼鼠白血病病毒核心以及两性鼠白血病病毒(FLYA13系)或猫内源性病毒RD114(FLYRD18系)的包膜糖蛋白。与以前的逆转录病毒包装系统的直接比较表明,FLYA13和FLYRD18系释放的滴度为无辅助重组病毒的100倍高。

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