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非缺陷型腺病毒2-猴病毒40杂交病毒的研究。3. Ad2+ND基因组的碱基组成、分子量和构象。

Studies of nondefective adenovirus 2-simian virus 40 hybrid viruses. 3. Base composition, molecular weight, and conformation of the Ad2+ND genome.

作者信息

Crumpacker C S, Henry P H, Kakefuda T, Rowe W P, Levin M J, Lewis A M

出版信息

J Virol. 1971 Mar;7(3):352-8. doi: 10.1128/JVI.7.3.352-358.1971.

Abstract

The nondefective adenovirus 2 (Ad2)-simian virus 40 (SV40) hybrid virus, Ad2(+)ND(1), differs from the defective Ad-SV40 hybrid populations previously described, in that this hybrid virus can replicate without the aid of nonhybrid adenovirus helper. Consequently, the deoxyribonucleic acid (DNA) from this virus, which can be obtained free of nonhybrid adenovirus DNA, is well suited for biophysical studies on Ad-SV40 hybrid DNA. Such studies have been performed and demonstrate Ad2(+)ND(1) DNA to have a buoyant density (1.715 g/cm(3)) and thermal denaturation profile (T(m) = 75.1 C) almost identical with nonhybrid Ad2 DNA. Furthermore, its molecular weight, as determined by analytical zone sedimentation and electron microscopy, was 22 x 10(6) to 25 x 10(6) daltons, which is also very similar to that determined for Ad2. Electron micrographs showed all of the hybrid molecules to be double-stranded and linear. By using this determination of the molecular weight of Ad2(+)ND(1) DNA and assuming that 1% of this molecule consists of covalently linked SV40 DNA (see companion paper), we calculate that the hybrid DNA molecule contains 220 x 10(3) to 250 x 10(3) daltons of SV40 DNA, or the equivalent of one-tenth of the SV40 genome.

摘要

无缺陷腺病毒2(Ad2)-猴病毒40(SV40)杂交病毒Ad2(+)ND(1),与先前描述的缺陷型Ad-SV40杂交群体不同,在于这种杂交病毒能够在没有非杂交腺病毒辅助的情况下进行复制。因此,这种病毒的脱氧核糖核酸(DNA)可以在不含非杂交腺病毒DNA的情况下获得,非常适合用于对Ad-SV40杂交DNA进行生物物理研究。已经进行了此类研究,结果表明Ad2(+)ND(1) DNA的浮力密度(1.715 g/cm³)和热变性曲线(T(m)=75.1℃)几乎与非杂交Ad2 DNA相同。此外,通过分析区带沉降和电子显微镜测定,其分子量为22×10⁶至25×10⁶道尔顿,这也与Ad2的测定值非常相似。电子显微镜照片显示所有杂交分子均为双链线性。通过对Ad2(+)ND(1) DNA分子量的这种测定,并假设该分子的1%由共价连接的SV40 DNA组成(见配套论文),我们计算出杂交DNA分子包含220×10³至250×10³道尔顿的SV40 DNA,相当于SV40基因组的十分之一。

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Conditional lethal mutants of adenovirus 2-simian virus 40 hybrids. I. Host range mutants of Ad2+ND1.
J Virol. 1974 Jun;13(6):1237-44. doi: 10.1128/JVI.13.6.1237-1244.1974.
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Use of nondefective adenovirus-simian virus 40 hybrids for mapping the simian virus 40 genome.
J Virol. 1973 Sep;12(3):643-52. doi: 10.1128/JVI.12.3.643-652.1973.
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Evidence for a transcription-control region of Simian virus 40 in the adenovirus 2--Simian virus 40 hybrid, Ad2+ND 1 .
Proc Natl Acad Sci U S A. 1972 Nov;69(11):3375-9. doi: 10.1073/pnas.69.11.3375.

本文引用的文献

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