Molecular Biology

核心A的总体目标是通过以下方式为计划项目提供分子生物学专业知识和支持 与所有三个项目和核心密切互动。核心A将构建新的猿人 编码HIV C株R5 env基因的免疫缺陷病毒(Shiv)株(简称 SHIVenvC‘s)；从感染SHIVenvC的猴子中分离、鉴定和测序env基因 疾病进展，并执行诊断性聚合酶链式反应以评估病毒载量。具体目标是： 1.将来自快速发展的艾滋病毒C分支疾病的赞比亚婴儿的env基因插入 倍增工程SIVmac239衍生的骨干，以增加复制能力 嵌合SHIVenvC菌株并加速灵长类动物的疾病进展。 2.为了基于不同于SIVmac239的SIV主干构建完全异源的SIV， 例如SIVsmE543-3，并通过插入与过去不同R5 HIV分支C膜基因 构建当前的SHIVenvC分离株。 3.产生和鉴定艾滋病毒分支C-env突变株，并与项目1密切合作，以评估 SHIVenvC感染恒河猴疾病进展过程中env的分子进化。 4.确保所有实验猴子在注册前没有其他逆转录病毒感染。一个 高灵敏的DMA聚合酶链式反应将被用来监测猴的D型逆转录病毒(SRV/D)或 猿猴嗜IV型病毒1型(STLV-1)。 5.用实时定量RT-PCR和DNA-PCR监测核心C区所有实验动物的血浆病毒 RNA或前病毒DMA分别加载。

The overall goal of Core A is to provide molecular biology expertise and support for the Program Project by closely interacting with all three Projects and Cores. Core A will construct novel simian-human immunodeficiency virus (SHIV) strains encoding R5 env genes of primary HIV clade C strains (termed SHIVenvC's); isolate, characterize, and sequence env genes from SHIVenvC-infected monkeys during disease progression, and perform diagnostic PCR to assess viral loads. The Specific Aims are: 1. To insert env genes derived from Zambian infants with rapidly progressive HIV clade C disease into multiply engineered SIVmac239-derived backbones to increase the replicative capacity of the chimeric SHIVenvC strains and to accelerate disease progression in primates. 2. To construct a fully heterologous SHIV based upon an SIV backbone that differs from SIVmac239, such as SIVsmE543-3, and by inserting an R5 HIV clade C env gene that differs from those used to construct the current SHIVenvC isolates. 3. To generate and characterize HIV clade C env mutants and to work closely with Project 1 to assess the molecular evolution of env in SHIVenvC-infected rhesus monkeys during disease progression. 4. To ensure that all experimental monkeys are free of other retrovirus infections prior to enrollment. A highly sensitive DMA PCR assay will be used to monitor for simian retrovirus type D (SRV/D) or simian Ivmphotropic virus type 1 (STLV-1). 5. To monitor all experimental animals in Core C by real-time RT-PCR and DNA PCR for plasma viral RNA or proviral DMA loads, respectively.