NATURE OF HIV-RELATED GENETIC SEQUENCES IN HUMAN DNA

采用灵敏的杂交和分子克隆技术， 允许鉴定和分离表现为 与所用探针的同源性低至60-70%，我们已经鉴定了新的 家族的远亲内源性逆转录病毒在各种 从鸟类到灵长类的脊椎动物物种，包括人类。 新的内源性逆转录病毒的鉴定，这些 程序已经揭示了逆转录病毒相关基因的存在， 在真核生物中单独存在的序列 基因组推测为反转录转座子或严重缺失的内源性 前病毒 利用类似的杂交技术，我们最近 鉴定的核苷酸序列显示与HIV相关基因的同源性， 未受感染的人类DNA序列。 因为这些艾滋病毒的存在- 未受感染的人类DNA中的相关基因序列， 这些序列对HIV的起源和致病性的影响， 我们建议：1）分离和鉴定这些HIV相关的遗传基因， 通过分子克隆和核苷酸分析， 分别进行排序，以明确其确切性质; 2)确定这些结构是否存在于其他脊椎动物中 属，试图确定它们的进化关系， 不同细胞类型中的相关结构，如果存在的话;和3）开始 识别和表征转录HIV相关遗传 如果它们确实可以被检测到的话。 如果这些与艾滋病有关的基因 结构可以通过核苷酸序列分析来证实， 进一步证明在人类细胞中表达，那么我们将 最终决定这些表达的生物学后果， 体内内源性HIV相关遗传结构，以确定 它们是否在HIV感染患者中发挥作用， 无症状、有症状或疾病的最终后果。 的 我们计划采用的实现这些目标的方法是 大部分已经在我们的实验室中实现，包括各种 物理化学、酶学和分子生物学技术 可用于DNA、RNA和蛋白质的分析。

Employing sensitive hybridization and molecular cloning techniques that allow identification and isolation of nucleotide sequences exhibiting as little as 60-70% homology to the probes employed, we have identified novel families of distantly related endogenous retroviruses in a variety of vertebrate species from avian to primate, including man. In addition to the identification of novel ancient endogenous retroviruses, these procedures have revealed the presence of retrovirus-related genetic sequences that are present individually (solo) throughout the eukaryotic genome presumably as retrotranspons or severely deleted endogenous proviruses. Utilizing similar hybridization techniques, we have recently identified nucleotide sequences exhibiting homology to HIV-related genetic sequences in uninfected human DNA. Because of the presence of these HIV- related genetic sequences in uninfected human DNA and the potential ramifications of these sequences to the origins and pathogenicity of HIV, we propose to: 1) isolate and characterize these HIV-related genetic structures present in human DNA by molecular cloning and nucleotide sequencing, respectively, in an effort to delineate their precise nature; 2) determine whether these structures are present in other vertebrate genera in an attempt to determine their evolutionary relationships with related structures in disparate cell types, if present; and 3) begin to identify and characterize the transcriptional HIV-related genetic structures if indeed they can be detected. If these HIV-related genetic structures can be substantiated by nucleotide sequence analysis and can further be demonstrated to be expressed in human cell, then we would ultimately determine the biological consequences of the expression of these endogenous HIV-related genetic structures in vivo in an effort to determine whether they play a role in HIV-infection in patients exhibiting asymptomatic, symptomatic, or terminal consequences of disease. THe methodologies that we plan to employ to achieve these objectives are for the most part already implemented in our laboratory and include a variety of physicochemical, enzymological, and molecular biological techniques available for the analysis of DNA, RNA, and protein.