CORE--RECOMBINANT DNA

核心——重组DNA

• 批准号: 6410446
• 负责人: ARTHUR ARMONE
• 金额: $ 17.7万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-01-01 至 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6410446
• 关键词: Escherichia coli; biomedical facility; gene expression; hemoglobin; human tissue; recombinant DNA; recombinant proteins; site directed mutagenesis

This core will create site-directed mutations of the alpha and beta globin subunits of human hemoglobin and will over-express these modified globins in E. coli using protocols that were optimized during last funding period of this Program Project Grant. In addition, strategies will be devised for producing globin subunits in which only a single type of amino acid is labeled with 13C. The frozen E. coli cell pasted produced in this core will be shipped to the Hemoglobin Assembly Core where the mutant hemoglobin tetramers will be assembled and purified.

该核心将创建人类血红蛋白的α和β球蛋白亚基的站点定向的突变，并使用在该计划项目赠款的最后资助期间优化的方案中过度表达这些改良的全球蛋白。此外，将制定生产球蛋白亚基的策略，其中只有单一类型的氨基酸标记为13C。该核心产生的粘贴的冷冻大肠杆菌细胞将运送到血红蛋白组件核心，其中突变的血红蛋白四聚体将组装并纯化。