HEPATITIS C--MODELS FOR REPLICATION

丙型肝炎病毒(HCV)是慢性肝炎的主要原因， 世界各地的肝功能衰竭和癌症并发症。前瞻 开发一种有效的疫苗仍然不太可能，而开发 新的抗病毒药物由于缺乏模型丙型肝炎病毒系统而受到限制。这个 部分慢性阻塞性肺疾病患者群体的进一步特征 以及研究丙型肝炎病毒复制所需的模型系统的开发 并以分子为基础的治疗学模型的确定和发展 研究丙型肝炎病毒复制和识别以分子为基础的 治疗学代表着尚未解决的主要问题。丙型肝炎病毒RNA依赖的RNA 聚合酶(RDRP)可能是丙型肝炎病毒抗病毒药物的靶点。我们已经表达了 重组丙型肝炎病毒RDRP是一种具有活性的酶。此外，我们 在一些人中检测到了重组丙型肝炎病毒RDRP的循环抗体 患者--这可能提供病毒复制的另一个指标。此外， 我们在开发一种在体内传播丙型肝炎病毒的系统方面取得了进展 培养的细胞。需要检验的假设是：体外模型和 可以开发细胞系统来研究丙型肝炎病毒复制的关键方面， 并确定直接针对丙型肝炎病毒的分子疗法 依赖于RNA的RNA聚合酶。 具体目标： 1)重组丙型肝炎病毒RNA依赖的RNA聚合酶的纯化和鉴定 (RDRP)，我们在大肠杆菌中表达，并在有活性的 表格；2)使用我们的丙型肝炎病毒RDRP分析来鉴定 直接抑制这种酶。3.使用纯化的丙型肝炎病毒RDRP进行 免疫分析将被用来进一步确定患者的特征 慢性丙型肝炎，以及4)为未来的水晶做可行性研究 增长是有效的。 利用表达活性的丙型肝炎病毒RNA=依赖的RA聚合酶的基因 开发模型细胞系统，使这方面的进一步研究成为可能 酶并识别进入完整细胞并抑制 丙型肝炎病毒RDRP。 继续研究一种有前途的传播丙型肝炎病毒的细胞培养系统 老鼠研究。我们的长期目标将是利用这个系统来研究 病毒基因表达和复制的细节，并识别移动的丙型肝炎病毒 抗病毒药物。

Hepatitis C virus (HCV) is a major cause of chronic hepatitis, with complications of liver failure and cancer, throughout the world. Prospects for developing an effective vaccine remain unlikely, and the development of new antivirals has been limited by the lack of model HCV systems. The further characterization of selected populations of patients with chronic HCV, and the development of model systems needed to study HCV replication and to identify molecular-based therapeutics and the development of model systems needed to study HCV replication and to identify molecular-based therapeutics represents major unsolved problems. The HCV RNA-dependent RNA polymerase (RDRP) is a likely target for HCV antivirals. We have expressed recombinant HCV RDRP an isolated it as an active enzyme. In addition, we have detected circulating antibodies to recombinant HCV RDRP in some patients - this may provide another index of viral replication. Moreover, we have made progress in developing a system that propagates HCV in cultured cells. The hypothesis to be tested is: that model in vitro and cellular systems can be developed to study key aspects of HCV replication, and to identify molecular-based therapeutics that directly target the HCV RNA-dependent RNA polymerase. Specific Objectives: 1) to purify and characterize recombinant HCV RNA-dependent RNA polymerase (RDRP) that we are expressing in E. coli and have isolated in an active form; 2) to use our HCV RDRP assay to identify small molecules that directly inhibit this enzyme. 3. To use purified HCV RDRP to perform immunoassays that will be used to further characterize patients with chronic hepatitis C, and 4) to do feasibility studies for future crystal growth work. The use the gene that expresses active HCV RNA=dependent RA polymerase to develop model cellular systems that will permit further studies of this enzyme and identify small molecules that enter intact cells and inhibit the HCV RDRP. To continue studies on a promising cell culture system that propagates HCV mouse studies. Our long-range goal will be to use this system to study details of viral gene expression and replication, and to identify moved HCV antiviral agents.