Department of Virology, Institute of Medical Microbiology, University of the Saarland, Homburg/Saar, Germany.
PLoS One. 2012;7(5):e36246. doi: 10.1371/journal.pone.0036246. Epub 2012 May 4.
In the nucleus of HIV-1 infected cells, unintegrated HIV-1 DNA molecules exist in the form of one and two LTR circles and linear molecules with degraded extremities. In tissue culture they are invariably more numerous than the provirus, the relative proportion of integrated to unintegrated forms varies widely from ∼1:1 to 1:10 and even over 1:100. In vivo, this ratio is unknown. To determine it, single nuclei from two infected patients with a known provirus copy number were microdissected, HIV DNA was amplified by nested PCR, cloned and individual clones sequenced. Given the extraordinary sequence complexity, we made the assumption that the total number of distinct sequences approximated to real number of amplifiable HIV-1 DNA templates in the nucleus. We found that the number of unintegrated DNA molecules increased linearly with the proviral copy number there being on average 86 unintegrated molecules per provirus.
在感染 HIV-1 的细胞的核内，未整合的 HIV-1 DNA 分子以一个和两个 LTR 环以及具有降解末端的线性分子的形式存在。在组织培养中，它们的数量总是比前病毒多，整合形式与未整合形式的相对比例从大约 1:1 到 1:10 甚至超过 1:100 不等。在体内，这个比例是未知的。为了确定这一点，从两名已知前病毒拷贝数的感染患者的单个核中进行显微切割，通过巢式 PCR 扩增 HIV DNA，克隆并对单个克隆进行测序。鉴于序列的复杂性，我们假设总序列数近似于核内可扩增 HIV-1 DNA 模板的实际数量。我们发现，未整合的 DNA 分子数量与前病毒拷贝数呈线性增加，平均每个前病毒有 86 个未整合的分子。
