A novel method to produce armored double-stranded DNA by encapsulation of MS2 viral capsids.
A novel method to produce armored double-stranded DNA by encapsulation of MS2 viral capsids.
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一种通过封装 MS2 病毒衣壳生产装甲双链 DNA 的新方法。
DOI：
10.1007/s00253-015-6664-4
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发表时间：
2015-09
影响因子：
5
通讯作者：
Li J
中科院分区：
文献类型：
--
作者：
Zhang L;Sun Y;Chang L;Jia T;Wang G;Zhang R;Zhang K;Li J
随着分子诊断技术的快速发展，越来越需要具有良好性能的质量控制和标准来监控整个检测过程。在这项研究中，我们描述了一种新的方法来生产装甲B型肝炎病毒（HBV）和人乳头瘤病毒（HPV）的DNA用于核酸检测，这被证实是稳定的，同质的，非感染性，耐核酸酶，和安全的运输。我们证明，MS 2噬菌体可以成功地包装双链DNA的1.3-，3-，3.5-，和6.5-kb的长度到病毒衣壳的重组效率高。这是RNA噬菌体MS 2作为包封双链DNA的平台的首次应用，形成在大小和形态上与天然MS 2衣壳难以区分的病毒样颗粒（VLP）。此外，通过分析pac位点与MS 2外壳蛋白（CP）的相互作用机制，我们发现除了识别的起始信号TR-RNA外，TR-DNA还可以触发CP二聚体的自发重组，为组装提供了一种更为方便可行的方法。总之，这种简单可靠的制备方法使装甲DNA成为临床实验室检测和诊断的理想对照品和标准品，具有广泛的应用前景，特别是为DNA病毒的质量控制提供了一个新的平台。本文的在线版本（doi：10.1007/s 00253 -015-6664-4）包含补充材料，可供授权用户使用。
With the rapid development of molecular diagnostic techniques, there is a growing need for quality controls and standards with favorable properties to monitor the entire detection process. In this study, we describe a novel method to produce armored hepatitis B virus (HBV) and human papillomavirus (HPV) DNA for use in nucleic acid tests, which was confirmed to be stable, homogeneous, noninfectious, nuclease resistant, and safe for shipping. We demonstrated that MS2 bacteriophage could successfully package double-stranded DNA of 1.3-, 3-, 3.5-, and 6.5-kb length into viral capsids with high reassembly efficiency. This is the first application of RNA bacteriophage MS2 as a platform to encapsulate double-stranded DNA, forming virus-like particles (VLPs) which were indistinguishable from native MS2 capsids in size and morphology. Moreover, by analyzing the interaction mechanism of pac site and the MS2 coat protein (CP), we found that in addition to the recognized initiation signal TR-RNA, TR-DNA can also trigger spontaneous reassembly of CP dimers, providing a more convenient and feasible method of assembly. In conclusion, this straightforward and reliable manufacturing approach makes armored DNA an ideal control and standard for use in clinical laboratory tests and diagnostics, possessing prospects for broad application, especially providing a new platform for the production of quality controls for DNA viruses. The online version of this article (doi:10.1007/s00253-015-6664-4) contains supplementary material, which is available to authorized users.
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影响因子：
17.1
作者：
Glasgow, Jeff E.;Capehart, Stacy L.;Francis, Matthew B.;Tullman-Ercek, Danielle
通讯作者：
Tullman-Ercek, Danielle
影响因子：
3.7
作者：
Obermeyer AC;Capehart SL;Jarman JB;Francis MB
通讯作者：
Francis MB
影响因子：
17.1
作者：
Ashley CE;Carnes EC;Phillips GK;Durfee PN;Buley MD;Lino CA;Padilla DP;Phillips B;Carter MB;Willman CL;Brinker CJ;Caldeira Jdo C;Chackerian B;Wharton W;Peabody DS
通讯作者：
Peabody DS
影响因子：
9.4
作者：
Konnick, EQ;Williams, SM;Hillyard, DR
通讯作者：
Hillyard, DR
影响因子：
9.4
作者：
Pasloske, BL;Walkerpeach, CR;DuBois, DB
通讯作者：
DuBois, DB