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利用重组痘苗病毒高效表达功能性腺病毒DNA聚合酶和前体末端蛋白。

High expression of functional adenovirus DNA polymerase and precursor terminal protein using recombinant vaccinia virus.

作者信息

Stunnenberg H G, Lange H, Philipson L, van Miltenburg R T, van der Vliet P C

机构信息

European Molecular Biology Laboratory, Heidelberg FRG.

出版信息

Nucleic Acids Res. 1988 Mar 25;16(6):2431-44. doi: 10.1093/nar/16.6.2431.

DOI:10.1093/nar/16.6.2431
PMID:3362670
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC336381/
Abstract

Initiation of Adenovirus (Ad) DNA replication occurs by a protein-priming mechanism in which the viral precursor terminal protein (pTP) and DNA polymerase (pol) as well as two nuclear DNA-binding proteins from uninfected HeLa cells are required. Biochemical studies on the pTP and DNA polymerase proteins separately have been hampered due to their low abundance and their presence as a pTP-pol complex in Ad infected cells. We have constructed a genomic sequence containing the large open reading frame from the Ad5 pol gene to which 9 basepairs from a putative exon were ligated. When inserted behind a modified late promoter of vaccinia virus the resulting recombinant virus produced enzymatically active 140 kDa Ad DNA polymerase. The same strategy was applied to express the 80 kDa pTP gene in a functional form. Both proteins were overexpressed at least 30-fold compared to extracts from Adenovirus infected cells and, when combined, were fully active for initiation in an in vitro Adenovirus DNA replication system.

摘要

腺病毒（Ad）DNA复制的起始通过蛋白质引发机制发生，该机制需要病毒前体末端蛋白（pTP）和DNA聚合酶（pol）以及来自未感染HeLa细胞的两种核DNA结合蛋白。由于pTP和DNA聚合酶蛋白丰度低且在Ad感染细胞中以pTP-pol复合物形式存在，分别对它们进行生化研究受到了阻碍。我们构建了一个基因组序列，其中包含来自Ad5 pol基因的大开放阅读框，并连接了来自一个推定外显子的9个碱基对。当插入痘苗病毒的修饰晚期启动子后面时，产生的重组病毒产生了具有酶活性的140 kDa Ad DNA聚合酶。同样的策略被用于以功能形式表达80 kDa pTP基因。与腺病毒感染细胞的提取物相比，这两种蛋白均过表达至少30倍，并且在体外腺病毒DNA复制系统中结合时，它们对起始反应具有完全活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f2a/336381/4806e1c8ec07/nar00149-0081-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f2a/336381/2141cd2307d9/nar00149-0079-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f2a/336381/4806e1c8ec07/nar00149-0081-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f2a/336381/2141cd2307d9/nar00149-0079-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f2a/336381/4806e1c8ec07/nar00149-0081-a.jpg

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本文引用的文献

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Cell. 1981 Feb;23(2):497-508. doi: 10.1016/0092-8674(81)90145-8.
2
Nucleotide sequences from the adenovirus-2 genome.来自腺病毒2型基因组的核苷酸序列。
J Biol Chem. 1982 Nov 25;257(22):13475-91.
3
Separation of the adenovirus terminal protein precursor from its associated DNA polymerase: role of both proteins in the initiation of adenovirus DNA replication.腺病毒末端蛋白前体与其相关DNA聚合酶的分离：两种蛋白在腺病毒DNA复制起始中的作用
通过对腺病毒 5 型 DNA 聚合酶进行合理的诱变，可以实现对腺病毒复制的选择性修饰。
J Virol. 2012 Oct;86(19):10484-93. doi: 10.1128/JVI.00739-12. Epub 2012 Jul 18.
4
Recruitment of the priming protein pTP and DNA binding occur by overlapping Oct-1 POU homeodomain surfaces.起始蛋白pTP的募集和DNA结合通过重叠的Oct-1 POU同源结构域表面发生。
EMBO J. 2002 Feb 15;21(4):725-35. doi: 10.1093/emboj/21.4.725.
5
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J Virol. 1999 Jun;73(6):4721-7. doi: 10.1128/JVI.73.6.4721-4727.1999.
6
The lipocalin Xlcpl1 expressed in the neural plate of Xenopus laevis embryos is a secreted retinaldehyde binding protein.在非洲爪蟾胚胎神经板中表达的脂质运载蛋白Xlcpl1是一种分泌型视黄醛结合蛋白。
Protein Sci. 1996 Jul;5(7):1250-60. doi: 10.1002/pro.5560050704.
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9
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10
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6
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8
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Curr Top Microbiol Immunol. 1984;109:53-73. doi: 10.1007/978-3-642-69460-8_2.