Hodge H M, Klein F, Bandyopadhyay A K, Robinson O R, Shibley G P
Appl Microbiol. 1974 Jan;27(1):224-31. doi: 10.1128/am.27.1.224-231.1974.
JLS-V9 mouse bone marrow cells were readily adapted to suspension culture, chronically infected with Rauscher leukemia virus (RLV), and subsequently grown in 7.5- and 14-liter New Brunswick fermentors. The suspension-type cell system can be modified to produce virus with clearly defined properties, such as high ribonucleic acid-dependent deoxyribonucleic acid polymerase (RDDP) activity, high particle count, and high infectious particle count. Biological and biophysical properties of suspension-produced RLV were not affected by concentration and purification employing continuous-flow and rate-zonal centrifugation procedures. The RDDP assay was standardized and showed a linear incorporation of (3)H-thymidine 5'-monophosphate ((3)H-TMP) up to 30 min. Further characterization indicated that a high percentage of (3)H-TMP incorporation was due to RDDP.
JLS-V9小鼠骨髓细胞易于适应悬浮培养,长期感染劳斯氏白血病病毒(RLV),随后在7.5升和14升的新布伦瑞克发酵罐中培养。悬浮型细胞系统可进行改良,以生产具有明确特性的病毒,如高核糖核酸依赖性脱氧核糖核酸聚合酶(RDDP)活性、高颗粒数和高感染性颗粒数。采用连续流和速率区带离心程序进行浓缩和纯化,不会影响悬浮培养产生的RLV的生物学和生物物理特性。对RDDP测定法进行了标准化,结果显示在长达30分钟的时间内,(3)H-胸苷5'-单磷酸((3)H-TMP)呈线性掺入。进一步的特性分析表明,(3)H-TMP的高掺入率归因于RDDP。