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潜伏感染的非洲绿猴肾细胞中的猿泡沫病毒3型(SFV-3):前病毒DNA去甲基化使其重新激活。

Simian foamy virus type 3 (SFV-3) in latently infected Vero cells: reactivation by demethylation of proviral DNA.

作者信息

Schweizer M, Fleps U, Jäckle A, Renne R, Turek R, Neumann-Haefelin D

机构信息

Abteilung Virologie, Universität Freiburg, Germany.

出版信息

Virology. 1993 Feb;192(2):663-6. doi: 10.1006/viro.1993.1084.

Abstract

Cell cultures latently infected with simian foamy virus type 3 (SFV-3) were established by suppressing lytic infection in Vero cells with 3'-azido-3'-deoxythymidine (AZT) and homologous antibodies (African green monkey serum immune to SFV-3). The resulting cell line, designated Vero-L, was shown to contain at least one copy per cell of SFV-3 DNA stably integrated at a defined site of the host cell genome. Sequencing of 669 bp at the integration site did not identify a coding region and revealed a 4-bp imperfect repeat in host cell DNA due to SFV-3 integration. Over 2 years of subcultivation, no spontaneous expression of proviral genes could be detected. However, the demethylating agent 5'-azacytidine reactivated lytic infection, proving conservation of the complete viral genome. Comparison of proviral DNA from latently and lytically infected cells supports the notion that methylation is instrumental in keeping SFV-3 infection in latency.

摘要

通过用3'-叠氮基-3'-脱氧胸苷(AZT)和同源抗体(对SFV-3免疫的非洲绿猴血清)抑制Vero细胞中的裂解感染,建立了潜伏感染猿猴泡沫病毒3型(SFV-3)的细胞培养物。所得细胞系命名为Vero-L,显示每个细胞至少含有一个稳定整合在宿主细胞基因组特定位点的SFV-3 DNA拷贝。对整合位点处669 bp的测序未鉴定出编码区,并揭示了由于SFV-3整合导致宿主细胞DNA中存在一个4 bp的不完全重复序列。在超过2年的传代培养中,未检测到前病毒基因的自发表达。然而,去甲基化剂5'-氮杂胞苷重新激活了裂解感染,证明了完整病毒基因组的保守性。潜伏感染和裂解感染细胞的前病毒DNA比较支持甲基化在使SFV-3感染保持潜伏状态中起作用的观点。

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