THE FUNCTION OF THE NEF PROTEIN OF SIV/MNE

SIV/MNE 的 NEF 蛋白的功能

• 批准号: 3874796
• 负责人: GISELA FANNING-HEIDECKER
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3874796
• 关键词: X ray crystallography; acquired immunodeficiency; gel electrophoresis; gene expression; gene induction /repression; genetic promoter element; insect virus; latent virus infection; monoclonal antibody; protein purification; protein structure function; serology /serodiagnosis; simian immunodeficiency virus; simian virus; transposon /insertion element; virus genetics; virus infection mechanism; virus protein

The function of the Nef protein encoded by HIV and simian immunodeficiency virus (SIV) is still unknown. Several reports propose that it is involved in viral latency due to a negative effect on viral transcription. Frequent inactivating mutations in the gene during virus isolation suggest that the gene is not tolerated well by tissue culture cells. In order to investigate the function of this gene we have generated several reagents. Expression of nef in the heterologous baculovirus expression heterologous system allows us to assess the serological status of animals infected with SIV/Mne. The protein is also purified by preparative gel electrophoresis and used to make poly- and monoclonal antibodies to replace a low titer rabbit antiserum that was made against a synthetic peptide. Furthermore, a mutant of the protein which is no longer myristylated is produced in large amounts for X-ray crystallography analysis. Nef expression constructs producing the gene product at constitutive high levels or under the control of a heat-inducible promoter have been generated to analyze the role of the Nef protein in infected and uninfected cells.

HIV编码的Nef蛋白的功能与猴免疫缺陷 病毒（SIV）仍然是未知的。几份报告提出， 在病毒潜伏期中，由于对病毒转录的负面影响。频繁 病毒分离过程中基因的失活突变表明， 基因不能被组织培养细胞很好地耐受。为了研究 这个基因的功能，我们已经产生了几种试剂。表达 异源杆状病毒表达异源系统中的nef允许 我们评估了SIV/Mne感染动物的血清学状态。的 还通过制备凝胶电泳纯化蛋白质， 制备多克隆抗体和单克隆抗体以替代低滴度的兔 抗合成肽的抗血清。此外，一个突变体 大量生产不再是肉豆蔻基化的蛋白质 用于X射线晶体学分析。Nef表达构建体，其产生 基因产物在组成型高水平或控制下， 已经产生了热诱导型启动子来分析Nef的作用， 感染和未感染细胞中的蛋白质。