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Efficient encapsidation of human immunodeficiency virus type 1 vectors and further characterization of cis elements required for encapsidation.

机译:人免疫缺陷病毒1型载体的有效衣壳化和衣壳化所需的顺式元件的进一步表征。

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摘要

To determine whether there is a cis-acting effect of translational expression of gag on RNA encapsidation, we compared the encapsidation of wild-type RNA with that of a mutant in which the translation of gag was ablated. This comparison indicated that there is not such a cis effect. To determine what is necessary and sufficient for encapsidation, we measured the relative encapsidation efficiencies of human immunodeficiency virus type 1 vector RNAs containing mutations in domains proximal to the canonical encapsidation signal or containing large deletions in the remainder of the genome. These data indicate that TAR and two additional regions are required for encapsidation and that the 5' end of the genome is sufficient for encapsidation. The Rev-responsive element is required mainly for efficient RNA transport from the nucleus to the cytoplasm. A foreign sequence was found to have a negative effect on encapsidation upon placement within the parental vector. Interestingly, this negative effect was compounded by multiple copies of the sequence.
机译:为了确定gag的翻译表达对RNA衣壳是否有顺式作用,我们将野生型RNA的衣壳与消去了gag的翻译突变体的衣壳进行了比较。该比较表明没有这种顺式作用。为了确定进行衣壳化的必要性和充分性,我们测量了人类免疫缺陷病毒1型载体RNA的相对衣壳化效率,该RNA在标准衣壳化信号附近的结构域中包含突变,或者在基因组的其余部分中包含大的缺失。这些数据表明,衣壳化需要TAR和另外两个区域,而基因组的5'端足以进行衣壳化。 Rev响应元件主要是RNA从细胞核到细胞质的有效转运所必需的。发现外源序列在亲本载体中放置后对衣壳化具有负面影响。有趣的是,这种负面影响由于该序列的多个拷贝而更加严重。

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