Martinez I, Dornburg R
Department of Molecular Genetics and Microbiology, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway 08854, USA.
Virology. 1995 Apr 1;208(1):234-41. doi: 10.1006/viro.1995.1147.
Using highly efficient gene expression vectors, we constructed new retroviral packaging lines derived from spleen necrosis virus. Core proteins are expressed from the murine leukemia virus promoter and enhancer followed by the tripartite leader sequence of an adenovirus. Using different plasmids for envelope expression, we found that the efficiency of vector transduction is dependent on the level of gag-pol expression. The level of envelope expression did not have a measurable impact on vector virus titers. The new helper cell lines do not contain any sequences homologous to vector genomes. They transduce standard retrovirus vectors with titers up to 10(6) colony forming units per milliliter of supernatant tissue culture medium. No replication-competent virus was observed.
我们使用高效基因表达载体构建了源自脾坏死病毒的新型逆转录病毒包装细胞系。核心蛋白由鼠白血病病毒启动子和增强子表达，随后是腺病毒的三联前导序列。使用不同质粒进行包膜表达时，我们发现载体转导效率取决于gag-pol的表达水平。包膜表达水平对载体病毒滴度没有可测量的影响。新的辅助细胞系不包含任何与载体基因组同源的序列。它们能转导标准逆转录病毒载体，上清组织培养基每毫升的滴度高达10(6)集落形成单位。未观察到有复制能力的病毒。
