EXPRESSION VECTOR ADMINISTRATION BY FEEDING R-ACNPV

通过饲喂 R-ACNPV 进行表达载体管理

• 批准号: 3498971
• 负责人: WANJUN LI
• 金额: $ 5万
• 依托单位: BIOKWITECH, INC.
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-05-15 至 1993-10-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3498971
• 关键词: Baculoviridae; Lepidoptera; SDS polyacrylamide gel electrophoresis; affinity chromatography; hemolymph; hepatitis B antigens; host organism interaction; inclusion body; insect virus; ion exchange chromatography; larva; method development; molecular biology; molecular cloning; nutrient intake activity; plaque assay; protein biosynthesis; protein purification; recombinant DNA; recombinant proteins; tissue /cell culture; transfection; transfection /expression vector; western blottings

Baculovirus protein expression in insect larvae provides a very useful, convenient, less labor-intensive and cost-effective system for the high- level production of proteins which may require a higher eukaryotic milieu for appropriate folding and/or post-translational modifications for biological activity. These in larva-produced proteins can be effectively used as veterinary diagnostics, therapeutics, vaccines, drug development, pesticides and antibody development. In our previous studies, we have been able to express various proteins in larvae by injection of recombinant baculoviruses. However, for the large-scale production of proteins in commercial markets, the efficiency of this system will be further improved by feeding the larvae with recombinant viruses instead of injection, as we have constructed a series of transfer plasmids for this purpose. In this project, we will set up this system and test the feasibility of this system for production of HBsAg in T. ni larvae with four specific aims: (1). to construct a recombinant transfer vector containing HBsAg; (2). to construct a recombinant AcNPV for larval feeding; (3). to feed larvae with recombinant AcNPV for production of HBsAg in T. ni larvae and (4). to purify HBsAg from haemolymph of infected larvae.

昆虫幼虫中的杆状病毒蛋白表达提供了非常有用的 方便，劳动密集型和成本效益的系统 蛋白质的水平生产可能需要更高的真核化学 环境适当的折叠和/或翻译后修改 用于生物活性。 这些在幼虫生产的蛋白质中可以是 有效用作兽医诊断，治疗剂，疫苗，药物 开发，农药和抗体开发。 在我们的上一个 研究，我们能够通过 注射重组杆菌病毒。 但是，对于大型 在商业市场中生产蛋白质，这是这样的效率 通过重组幼虫进食幼虫将进一步改善系统 病毒而不是注射，因为我们已经构建了一系列 为此目的转移质粒。 在这个项目中，我们将设置 该系统并测试该系统生产的可行性 T. ni幼虫中的Hbsag具有四个具体目标：（1）。构造一个 重组转移载体含有HBSAG； （2）。构造一个 重组ACNPV用于幼虫进食； （3）。用 重组ACNPV用于在T. ni幼虫和（4）中生产HbSAg。到 从感染幼虫的血液中纯化Hbsag。