Cotten M, Saltik M, Kursa M, Wagner E, Maass G, Birnstiel M L
Research Institute of Molecular Pathology, Wien, Austria.
Virology. 1994 Nov 15;205(1):254-61. doi: 10.1006/viro.1994.1641.
Adenovirus entry into its host cell transiently permeabilizes the cell allowing the coentry of reagents such as DNA. We compare here adenovirus inactivation with beta-propiolactone and several psoralen derivatives, seeking reagents that disrupt the viral genome without impairing the viral entry functions. No virus replication can be detected after 8-methoxypsoralen (8-MOP) modification. Viral transcription is not detectable by Northern analysis, and reverse transcriptase/PCR analysis demonstrates at least a 1000-fold decrease in viral transcription after 8-MOP treatment. Using [3H]8-MOP, the psoralen is found to enter the virus capsid and react throughout the viral genome, with approximately one psoralen modification per 100 bp of viral DNA. This inactivated adenovirus allows us to deliver DNA to target cells without interference from adenovirus gene expression or replication. Furthermore, we can now study the host cell response to adenovirus entry without the complications of adenovirus gene expression.
腺病毒进入宿主细胞会使细胞短暂通透，从而允许诸如DNA等试剂共同进入。我们在此比较腺病毒用β-丙内酯和几种补骨脂素衍生物进行灭活的情况，寻找能破坏病毒基因组而不损害病毒进入功能的试剂。用8-甲氧基补骨脂素（8-MOP）处理后未检测到病毒复制。通过Northern分析未检测到病毒转录，逆转录酶/聚合酶链反应分析表明，8-MOP处理后病毒转录至少下降1000倍。使用[3H]8-MOP，发现补骨脂素进入病毒衣壳并与整个病毒基因组发生反应，每100个碱基对的病毒DNA约有一个补骨脂素修饰。这种灭活的腺病毒使我们能够将DNA递送至靶细胞，而不受腺病毒基因表达或复制的干扰。此外，我们现在可以研究宿主细胞对腺病毒进入的反应，而不会有腺病毒基因表达带来的复杂情况。
