DELETION MUTAGENESIS WITHIN NONCODING SEGMENTS OF HIV1

描述：这项建议的长期目标是更好地理解 HIV的非编码区，以阐明这一贡献 反转录、病毒基因表达、蛋白质合成的区域 和病毒复制。该区域的缺失突变也可能有助于 开发一种减毒艾滋病毒疫苗，并广泛研究使用 将进行SIV模型，以确定是否可能是这种情况。 这项研究将提供关于相对较差的 了解艾滋病毒基因组的区域。在需要回答的问题中，有 如下：1.是否删除下游的54个核苷酸片段 引子结合部位将永久减弱HIV在组织中的复制 培养，并了解涉及一种 54个核苷酸区域内的一系列较小的缺失。要确定是否存在 54nT区3‘端的缺失突变也可能危害HIV 复制能力并干扰病毒DNA、mRNA、 和蛋白质，其程度与54个核苷酸缺失的程度相同。2.确定 猴免疫缺陷病毒(SIV)中是否存在类似的缺失 在HIV的54个核苷酸缺失中，也会对病毒产生严重影响 复制，病毒DNA的产生，病毒mRNA的合成，以及 生产病毒蛋白。3.是否将54个核苷酸的缺失与 先前被显示为提高RT保真度的RT中的184V替换可以 减少发生逆转的可能性。另外，这将是什么 RT中的184V替换与删除 富于循环的还原速度？4.判断是否 SIV中大小缺失突变的组合，以及 RT中184V的替代提高了保真度，同样会导致 在较长时间内回复到野生型病毒的可能性较小。 5.确定SIV变异体，包括在 在RT中具有或不具有184V替换的非编码区可以作为 用于刺激抗SIV体液和 猕猴的细胞免疫反应性。6.确定这些项目是否 衰减的SIV构造可以防止后续的挑战 毒力形式的SIV和SIV/HIV杂交体。

DESCRIPTION: The long term goal of this proposal is to better understand the non-coding region of HIV so as to shed light on the contribution of this region to reverse transcription, viral gene expression, protein synthesis and viral replication. Deletion mutagenesis in this region may also help in the development of an attenuated HIV vaccine, and extensive research using the SIV model will be conducted to determine whether this may be the case. This research will provide important new information on a relatively poorly understood area of the HIV genome. Among the questions to answer are the following: 1. Whether a deletion of the 54 nt segment downstream of the primer binding site will permanently attenuate HIV replication in tissue culture, and to understand the molecular basis for reversions that involve a series of smaller deletions within the 54 nt region. To determine whether a deletion mutation at the 3' end of the 54 nt region may also compromise HIV replication capacity and interfere with the production of viral DNA, mRNA, and proteins to the same extent as the 54 nt deletion. 2. To determine whether a similar deletion in simian immunodeficiency virus (SIV), to that of the 54 nt deletion in HIV, will also impact severely on viral replication, generation of viral DNA, synthesis of viral mRNA, and production of viral protein. 3. Whether combining the 54 nt deletion with the 184V substitution in RT, previously shown to increase RT fidelity, may diminish the likelihood of reversion taking place. Also, what will the effect be of combining the 184V substitution in RT with deletion of the A-rich loop on the rapidity of reversion? 4. To determine whether combination of the large and small deletion mutations in SIV, together with the 184V substitution in RT that increases fidelity, will likewise result in a lesser likelihood of reversion to wild-type virus over extended periods. 5. To determine whether SIV variants, containing large deletions in the non-coding region, with or without the 184V substitution in RT, may serve as attenuated vaccines for purposes of stimulating anti-SIV humoral and cellular immune responsiveness in macaques. 6. To determine whether these attenuated SIV constructs may protect against subsequent challenge with virulent forms of SIV and SIV/HIV hybrids.