Particle infectivity of HIV-1 full-length genome infectious molecular clones in a subtype C heterosexual transmission pair following high fidelity amplification and unbiased cloning.
Particle infectivity of HIV-1 full-length genome infectious molecular clones in a subtype C heterosexual transmission pair following high fidelity amplification and unbiased cloning.
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DOI：
10.1016/j.virol.2014.08.018
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发表时间：
2014-11
期刊：
影响因子：
3.7
通讯作者：
Hunter, Eric
中科院分区：
文献类型：
--
作者：
Deymier, Martin J.;Claiborne, Daniel T.;Ende, Zachary;Ratner, Hannah K.;Kilembe, William;Allen, Susan;Hunter, Eric
HIV-1的高遗传多样性阻碍了常用限制性内切酶方法的高通量、大规模测序和全长基因组克隆。利用高保真聚合酶进行扩增和无偏融合克隆策略的新方法，我们从流行病学上相关的传播对中生成了几个HIV-1全长基因组感染性分子克隆。这些克隆代表了传播/创始人病毒和来自慢性感染个体在传播时间附近的不同准种的系统发育多样性的非传播变体。我们证明，使用这种方法，同源单基因组扩增子衍生的全长克隆中pcr诱导的突变是罕见的。此外，测试的所有8个非传播基因组都产生了具有一系列传染性的功能性病毒，这掩盖了之前的假设，即慢性HIV-1感染中的大多数循环病毒是有缺陷的。因此，这些方法为分子生物学方案的更新提供了重要的工具，可以普遍应用于人类病毒病原体的研究。
The high genetic diversity of HIV-1 impedes high throughput, large-scale sequencing and full-length genome cloning by common restriction enzyme based methods. Applying novel methods that employ a high-fidelity polymerase for amplification and an unbiased fusion-based cloning strategy, we have generated several HIV-1 full-length genome infectious molecular clones from an epidemiologically linked transmission pair. These clones represent the transmitted/founder virus and phylogenetically diverse non-transmitted variants from the chronically infected individual's diverse quasispecies near the time of transmission. We demonstrate that, using this approach, PCR-induced mutations in full-length clones derived from their cognate single genome amplicons are rare. Furthermore, all eight non-transmitted genomes tested produced functional virus with a range of infectivities, belying the previous assumption that a majority of circulating viruses in chronic HIV-1 infection are defective. Thus, these methods provide important tools to update protocols in molecular biology that could be universally applied to the study of human viral pathogens.
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DOI：
10.1126/science.1254031
发表时间：
2014-07-11
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Carlson JM;Schaefer M;Monaco DC;Batorsky R;Claiborne DT;Prince J;Deymier MJ;Ende ZS;Klatt NR;DeZiel CE;Lin TH;Peng J;Seese AM;Shapiro R;Frater J;Ndung'u T;Tang J;Goepfert P;Gilmour J;Price MA;Kilembe W;Heckerman D;Goulder PJ;Allen TM;Allen S;Hunter E
通讯作者：
Hunter E
影响因子：
3.7
作者：
Hotard AL;Shaikh FY;Lee S;Yan D;Teng MN;Plemper RK;Crowe JE Jr;Moore ML
通讯作者：
Moore ML
DOI：
10.1073/pnas.1304288110
发表时间：
2013-04-23
影响因子：
11.1
作者：
Parrish, Nicholas F.;Gao, Feng;Hahn, Beatrice H.
通讯作者：
Hahn, Beatrice H.
影响因子：
3.7
作者：
Ndung'u, T;Renjifo, B;Essex, M
通讯作者：
Essex, M
DOI：
10.1073/pnas.0802203105
发表时间：
2008-05-27
影响因子：
11.1
作者：
Keele, Brandon F.;Giorgi, Elena E.;Shaw, George M.
通讯作者：
Shaw, George M.