腺相关病毒增殖。IX. 病毒基因组在体内的转录程度。
Adenovirus-associated virus multiplication. IX. Extent of transcription of the viral genome in vivo.
作者信息
Carter B J, Khoury G, Rose J A
出版信息
J Virol. 1972 Dec;10(6):1118-25. doi: 10.1128/JVI.10.6.1118-1125.1972.
Abstract
Nucleic acid hybridization procedures were used to measure the extent of transcription of adenovirus-associated virus (AAV) deoxyribonucleic acid (DNA) in KB cells in the presence of either adenovirus or herpes simplex virus as the helper. Annealing of AAV ribonucleic acid to AAV DNA was monitored by a hybridization inhibition assay on nitrocellulose filters or by hydroxyapatite chromatography. These experiments confirmed the previous observation that, in the presence of either type of helper virus, only one strand of AAV DNA (the thymidine-rich or "minus" strand) is transcribed in vivo. However, it was found that only 70 to 80% of this strand appears to be transcribed in vivo. Furthermore, studies with minus strands employing hydroxyapatite chromatography and nuclease S(1), which specifically degrades single-stranded DNA, indicated that up to 20% of the minus strand is self-complementary. It seems likely that these self-complementary sequences account for the bulk of that portion of the minus strand (20 to 30%) which is not transcribed in vivo.
摘要
利用核酸杂交程序来测定在腺病毒或单纯疱疹病毒作为辅助病毒存在的情况下，KB细胞中腺病毒相关病毒（AAV）脱氧核糖核酸（DNA）的转录程度。通过在硝酸纤维素滤膜上进行杂交抑制试验或通过羟基磷灰石层析来监测AAV核糖核酸与AAV DNA的退火情况。这些实验证实了先前的观察结果，即在任何一种辅助病毒存在的情况下，体内只有AAV DNA的一条链（富含胸腺嘧啶的或“负”链）被转录。然而，发现这条链在体内似乎只有70%至80%被转录。此外，利用羟基磷灰石层析和核酸酶S(1)（其特异性降解单链DNA）对负链进行的研究表明，高达20%的负链是自我互补的。似乎很可能这些自我互补序列占负链中未在体内转录的那部分（20%至30%）的大部分。
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