...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Virology >Synthesis of Virus-Specific Ribonucleic Acid in KB Cells Infected with Type 2 Adenovirus
【24h】

Synthesis of Virus-Specific Ribonucleic Acid in KB Cells Infected with Type 2 Adenovirus

机译：用2型腺病毒感染KB细胞中病毒特异性核糖核酸的合成

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

By using the technique of deoxyribonucleic acid (DNA)-ribonucleic acid (RNA) hybridization, virus-specific RNA (cRNA) was detected 6 hr after infection in preparations of total RNA from cells infected with type 2 adenovirus in the presence of 2 μm 5-fluorodeoxyuridine. In the absence of 5-fluorodeoxyuridine, there was a continuous increase in the incorporation of 3H-uridine into viral cRNA until 20 hr after infection, at which time approximately 40% of the 3H-uridine entering RNA was found in virus-specific RNA. When RNA was prepared from polyribosome fractions obtained from cytoplasmic extracts of infected cells, virus-directed transcription was detected at 3 hr after infection (i.e., 3 to 4 hr before the initiation of viral DNA synthesis). Viral cRNA species synthesized at different times after infection were compared by the technique of DNA-RNA hybridization-inhibition (“presaturation” hybridization-competition). Three hybridization-inhibition techniques were compared. The techniques differed in the manner in which the DNA-RNA complex was isolated after the first hybridization reaction. Depending on the procedure employed, various degrees of inhibition were measured. The variation could be essentially eliminated if prior to hybridization the inhibitory RNA species were alkali-degraded to a uniform size of about 4S. Undegraded RNA could be used if the DNA-RNA complex was isolated by using a procedure involving rigorous washing (preferably including ribonuclease treatment) before the second hybridization with labeled RNA. When a rigorous hybridization-inhibition procedure was used, three classes of virus-specific RNA species could be distinguished: (i) early RNA class I whose synthesis began prior to viral DNA replication and stopped at some time after the initiation of viral DNA replication—it comprised about 70% of the early RNA species and was apparently degraded by 18 hr after infection; (ii) early RNA class II whose synthesis began prior to viral DNA replication and apparently continued at an enhanced rate late in infection; and (iii) late RNA whose synthesis began after the initiation of viral DNA synthesis.
机译：通过使用脱氧核糖核酸（DNA） - 柠檬酸（RNA）杂交的技术，在感染后6小时检测到病毒特异性RNA（CRNA），在2μm5的情况下感染2型腺病毒的细胞中的总RNA的制剂 - 氟二氢脲酰亚胺。在没有5-氟脱氧核素尿苷的情况下，在感染后将 3-sup> H-尿苷掺入病毒CRNA至20小时的情况下连续增加，此时约40％的 3 < / sup> H-尿苷进入病毒特异性RNA中的RNA。当从受感染细胞的细胞质提取物中获得的多吡菊组馏分制备RNA时，在感染后3小时检测病毒的转录（即，在引发病毒DNA合成之前，3至4小时）。通过DNA-RNA杂交抑制技术（“最大”杂交 - 竞争）进行了在感染后在不同时间合成的病毒性CRNA物种。比较了三种杂交抑制技术。该技术以第一杂交反应在第一杂交反应后分离的方式不同。根据所用方法，测量各种抑制程度。如果在杂交之前，可以基本上消除变化，抑制RNA物种被碱 - 降解到约4 S 的均匀尺寸。如果通过使用涉及严格的洗涤（优选包括核糖核酸酶处理）之前的第二杂交在标记的RNA之前分离DNA-RNA复合物，则可以使用未解析的RNA。当使用严格的杂交抑制程序时，可以区分三类病毒特异性RNA物种：（i）早期的RNA类I，其合成在病毒DNA复制之前开始于病毒DNA复制并在发生病毒DNA复制后的一段时间停止，它包含约70％的早期RNA物种，感染后明显降解18小时; （ii）早期的RNA类II，其合成在病毒DNA复制之前开始，并且显然以感染后期增强的速率持续; （iii）合成在发生病毒DNA合成后开始的晚期RNA。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号