Mini Chromosome and Fluorescent Proteins for Expression of Protein Complexes

用于蛋白质复合物表达的微型染色体和荧光蛋白

• 批准号: 9026623
• 负责人: Ronald Godiska
• 金额: $ 76.7万
• 依托单位: LUCIGEN CORPORATION
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-21 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9026623
• 关键词: Acute; Angiogenesis Modulating Agents; Basic Science; Biochemical Pathway; Biological Assay; Cell Culture Techniques; Cell Line; Cells; Characteristics; Chromosomes; Cloning; Cloning Vectors; Complementary DNA; Complex; Computer Simulation; DNA Sequence; DNA cassette; Development; Elements; Energy Transfer; Enzymes; Episome; Escherichia coli; Excision; Extinction (Psychology); Feedback; Fluorescence; Funding; Gene Expression; Genes; Genetic; Goals; Growth; Human; Human Genome; Human body; In Vitro; Individual; Ion Channel; Knowledge; Libraries; Light; Mammalian Cell; Methods; Modeling; Monitor; Mutagenesis; Mutate; Mutation; Neurons; Operon; Pattern; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Process; Protein Analysis; Protein Complex Subunit; Proteins; Regulation; Reporter; Reporting; Research; Research Personnel; Scientific Advances and Accomplishments; Shuttle Vectors; Small Business Innovation Research Grant; Staging; System; Therapeutic; Transfection; Tumor Suppression; Validation; Variant; abstracting; arm; base; commercialization; design; expression cloning; expression vector; high throughput screening; improved; in vitro Model; in vivo; inducible gene expression; mRNA Decay; meetings; novel; promoter; protein complex; protein expression; protein protein interaction; quantum; research and development; stable cell line; synthetic biology; temporal measurement; tool; tumor; vector; viral DNA

Summary/abstract The proteins encoded by the human genome may participate in as many as 200,000 pairwise protein interactions, and “nearly every major process in a cell is carried out by assemblies of 10 or more interlocking protein molecules” (Bruce Alberts, Cell 1998). However, there are no commercially available expression systems for regulated expression of multi-subunit protein complexes. There is an acute need for a mammalian expression system that can produce multi-protein complexes, is tightly regulated, stable in prolonged culture, and non-integrated. No reported expression system comes near to meeting all these needs. Lucigen will develop and commercialize a stable episomal expression system with unbiased cloning ability, tunable light-inducible promoters, improved fluorescent tags, and the ability to express equivalent amounts of multiple proteins. This system will facilitate direct experimental validation of the myriad protein complexes predicted by computational models, as well as those that are already known (e.g, ion channels, multi-subunit enzymes, biochemical pathways).

摘要/摘要 人类基因组编码的蛋白质可能参与多达200,000个成对蛋白质 相互作用，以及细胞中几乎每一个主要过程都是由10人或更多人组成的组件进行的 互锁蛋白质分子“(Bruce Alberts，Cell 1998)。然而，目前还没有商业上可用的 多亚单位蛋白复合体调控表达的表达系统。有一种迫切的需求 对于能够产生多蛋白复合体的哺乳动物表达系统，受到严格调控， 长期培养稳定，无整合性。目前还没有报道的表达系统即将面世 所有这些需求。Lucigen将开发稳定的表观表达系统并将其商业化 无偏见的克隆能力，可调的光诱导启动子，改进的荧光标签，以及 表达等量的多种蛋白质。该系统将便于直接进行实验 验证由计算模型预测的无数蛋白质复合体，以及那些 已知的(例如，离子通道、多亚单位酶、生化途径)。