FACTOR(S) CONTROLLING GLOBIN GENE EXPRESSION IN K562 CELLS

K562 细胞中控制球蛋白基因表达的因子

• 批准号: 4689443
• 负责人: J ELION
• 金额: --
• 依托单位: NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES
• 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/4689443
• 关键词: azacitidine; gel filtration chromatography; gene expression; genes; genetic manipulation; genetic transcription; human tissue; ion exchange chromatography; messenger RNA; plasmids; tissue /cell culture

K562 cells are used as a model to study the control of human globin genes expression. They express embryonic and fetal but not adult globin genes. Results suggests that the lack of transcription of the adult beta-globin gene results from the absence/presence of an activating/repressing trans-acting activator or a repressor. This is being tested by studying the transient expression of the bacterial CAT enzyme in K562 cells transfected with plasmids in which the transcription of the CAT gene is driven by various globin gene promoters. In competition experiments, cells are co-transfected with the same plasmids and an excess of the corresponding globin promoter. Similar experiments are conducted with K562 cells grown in the presence of 5-azacytidine, which has been previously shown to induce beta-globin gene transcription in these cells. This represents a necessary step toward further characterization and isolation of this trans-acting factor(s).

以K562细胞为模型，研究人珠蛋白基因的调控 表情。它们表达胚胎和胎儿的珠蛋白基因，但不表达成体珠蛋白基因。 结果表明，成人β-珠蛋白的转录缺失 基因是由激活/抑制的缺失/存在引起的 反式激活因子或抑制因子。这一点正在通过研究进行检验 细菌CAT酶在K562细胞中的瞬时表达 用其中CAT基因的转录被 由各种珠蛋白基因启动子驱动。在竞争实验中，细胞 与相同的质粒共转染，并且过量的 相应的珠蛋白启动子。K562也进行了类似的实验 在5-氮胞苷存在下生长的细胞，以前已经 被证明能在这些细胞中诱导β-珠蛋白基因转录。这 代表了进一步描述和隔离的必要步骤 这个反式作用因素(S)。