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劳氏鼠白血病病毒的体外产生:培养时间对生物学特性的影响。

In vitro production of Rauscher murine leukemia virus: influence of culture age on biological properties.

作者信息

Robinson O R, Perry A, Shibley G P, de Maio M

出版信息

Appl Environ Microbiol. 1979 Dec;38(6):1132-9. doi: 10.1128/aem.38.6.1132-1139.1979.

Abstract

Large-scale production and concentration procedures have been standardized to study the biological properties of Rauscher leukemia virus produced from the high-passaged JLS-V9-H mouse bone marrow cell line. Virus produced early (days 4 to 6) in the harvest and refeed cycle contained higher levels of ribonucleic acid-directed deoxyribonucleic acid polymerase activity and was more infectious than Rauscher leukemia virus produced later (days 7 to 10) in the growth period. The peak of virus production as detected by physical assays (virus particle count, protein, and p30 antigen) was highest at day 6, whereas the optimum biological and ribonucleic acid-directed deoxyribonucleic acid polymerase activity occurred 24 h earlier. When product characterization values of each concentrate were adjusted to a specific activity (i.e., per milligram of protein) basis, virus particle counts averaged 4 x 10(11) through days 5 to 9, and the peak infectivity occurred at day 4, whereas ribonucleic acid-directed deoxyribonucleic acid polymerase activity was highest at day 4 (endogenous) and 5 (exogenous). Sodium dodecyl sulfate-polyacrylamide gel analysis revealed only slight differences in the polypeptide pattern of Rauscher leukemia virus harvested from cultures of varying age, although Rauscher leukemia virus produced between days 3 and 5 contained more glycoprotein than either earlier or later harvests.

摘要

为了研究从高传代JLS-V9-H小鼠骨髓细胞系产生的劳氏肉瘤病毒的生物学特性,大规模生产和浓缩程序已实现标准化。在收获和再投喂周期早期(第4至6天)产生的病毒含有更高水平的核糖核酸指导的脱氧核糖核酸聚合酶活性,并且比在生长后期(第7至10天)产生的劳氏肉瘤病毒更具传染性。通过物理测定(病毒颗粒计数、蛋白质和p30抗原)检测到的病毒产生高峰在第6天最高,而最佳生物学活性和核糖核酸指导的脱氧核糖核酸聚合酶活性则提前24小时出现。当将每种浓缩物的产品特性值调整到比活性(即每毫克蛋白质)基础时,在第5至9天病毒颗粒计数平均为4×10¹¹,峰值感染性出现在第4天,而核糖核酸指导的脱氧核糖核酸聚合酶活性在第4天(内源性)和第5天(外源性)最高。十二烷基硫酸钠-聚丙烯酰胺凝胶分析显示,从不同年龄培养物中收获的劳氏肉瘤病毒的多肽模式只有轻微差异,尽管在第3至5天产生的劳氏肉瘤病毒比早期或晚期收获的含有更多糖蛋白。

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