DELETION MUTAGENESIS WITHIN NONCODING SEGMENTS OF HIV1

描述：本提案的长期目标是更好地了解 艾滋病毒的非编码区，以阐明这一贡献， 逆转录区域，病毒基因表达，蛋白质合成 和病毒复制。 该区域的缺失突变也可能有助于 一种减毒艾滋病毒疫苗的开发，以及使用 将进行SIV模型以确定是否可能是这种情况。 这项研究将提供重要的新信息， 了解艾滋病毒基因组的区域。 要回答的问题包括 以下内容：1. 是否缺失54 nt片段下游的 引物结合位点将永久减弱组织中的HIV复制 文化，并了解分子基础的逆转，涉及一个 54 nt区域内的一系列较小缺失。 以确定是否 54 nt区域3'端的缺失突变也可能损害HIV 复制能力和干扰病毒DNA，mRNA， 以及与54 nt缺失相同程度的蛋白质。 2. 以确定 猿免疫缺陷病毒（SIV）中是否存在类似的缺失， HIV中54 nt缺失的基因，也将严重影响病毒的 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 nt缺失与 先前显示增加RT保真度RT中的184 V取代可 减少了逆转发生的可能性。 还有， 将RT中的184 V取代与RT中的184 V取代的缺失相结合， 一个丰富的循环对逆转的速度？ 4. 以确定是否 SIV中大和小缺失突变的组合，以及 RT中增加保真度的184 V取代同样会导致 在较长的时间内回复到野生型病毒的可能性较小。 5. 为了确定是否SIV变异，包含大的缺失， 在RT中具有或不具有184 V置换的非编码区可以充当 用于刺激抗SIV体液免疫和 猕猴的细胞免疫反应。 6. 以确定是否这些 减毒SIV构建体可以保护免受随后的攻击， SIV和SIV/HIV杂交体的毒性形式。

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.