Ban J, First N L, Temin H M
McArdle Laboratory for Cancer Research, University of Wisconsin, Madison 53706.
J Gen Virol. 1989 Aug;70 ( Pt 8):1987-93. doi: 10.1099/0022-1317-70-8-1987.
Retroviral packaging cell lines were constructed by using the gag-pol gene of spleen necrosis virus, the gag-pol gene of Moloney murine leukaemia virus and the env gene of bovine leukaemia virus. The plasmids containing the gag-pol genes and the plasmid containing the env gene were cotransfected into NIH/3T3 and D17 cells. The cells containing the helper virus constructs were tested for their ability to package replication-defective murine leukaemia and avian reticuloendotheliosis retrovirus vectors. The titre of vector virus produced by each of the retroviral packaging cell lines was about 10(2) colony-forming units per ml of medium. Tests for events that might result in intact replication-competent retroviruses showed no evidence for the generation of such viruses. The vector viruses were able to infect dog and rat cells. Bovine cells were infected only after their cocultivation with the retroviral packaging cell lines producing murine leukaemia virus vectors, perhaps as a result of a low concentration of receptors.
通过使用脾坏死病毒的gag-pol基因、莫洛尼鼠白血病病毒的gag-pol基因和牛白血病病毒的env基因构建逆转录病毒包装细胞系。将含有gag-pol基因的质粒和含有env基因的质粒共转染到NIH/3T3和D17细胞中。对含有辅助病毒构建体的细胞进行测试，以检测其包装复制缺陷型鼠白血病和禽网状内皮组织增生症逆转录病毒载体的能力。每个逆转录病毒包装细胞系产生的载体病毒滴度约为每毫升培养基10(2)个集落形成单位。对可能导致产生完整的具有复制能力的逆转录病毒的事件进行检测，未发现产生此类病毒的证据。载体病毒能够感染狗和大鼠细胞。牛细胞只有在与产生鼠白血病病毒载体的逆转录病毒包装细胞系共培养后才能被感染，这可能是由于受体浓度较低所致。
