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SV40 DNA对猴细胞的高效感染。II. 低分子量DEAE-葡聚糖在大规模实验中的应用。

Efficient infection of monkey cells with SV40 DNA. II. Use of low-molecular-weight DEAE-dextran for large-scale experiments.

作者信息

Danna K J, Sompayrac L M

出版信息

J Virol Methods. 1982 Dec;5(5-6):335-41. doi: 10.1016/0166-0934(82)90025-8.

Abstract

With standard protocols for DNA infection, only a small percentage (about 4%) of monkey ells exposed to purified DNA of simian virus 40 (SV40) displays cytopathic effect or expresses viral T antigen. We have recently reported (Sompayrac and Danna, 1981) that by extending the time of exposure of BSC-1 cells to DNA in the presence of low concentrations of diethylaminoethyl (DEAE)-dextran, we can infect up to 50% of the cells. Because our protocol was devised for DEAE-dextran of mol. wt. 2 X 10(6), which is no longer commercially available, we have tested a low-molecular-weight polymer (mol. wt. 0.5 X 10(6) that can be purchased. Use of this reagent in our protocol resulted in marginally higher levels of infection than we found with the high-molecular-weight polymer. However, slightly more DNA (about 1.5 times as much) was needed to achieve saturation. With both reagents, the percentage of cells infected was proportional to time of exposure. Therefore, low-molecular-weight DEAE-dextran should be useful for large-scale DNA infections.

摘要

按照标准的DNA感染方案,用猿猴病毒40(SV40)的纯化DNA处理猴细胞时,只有一小部分(约4%)细胞会出现细胞病变效应或表达病毒T抗原。我们最近报道过(Sompayrac和Danna,1981年),在低浓度二乙氨基乙基(DEAE)-葡聚糖存在的情况下,延长BSC-1细胞与DNA的接触时间,我们能够感染多达50%的细胞。由于我们的方案是针对分子量为2×10⁶的DEAE-葡聚糖设计的,而这种产品已不再有商业供应,我们测试了一种可以购买到的低分子量聚合物(分子量为0.5×10⁶)。在我们的方案中使用这种试剂,感染水平比使用高分子量聚合物时略高。然而,达到饱和需要略多一些的DNA(约为1.5倍)。使用这两种试剂时,被感染细胞的百分比与接触时间成正比。因此,低分子量的DEAE-葡聚糖应该可用于大规模的DNA感染。

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