The Knockout of the Mouse Muc5ac

小鼠 Muc5ac 的敲除

• 批准号: 7734559
• 负责人: Robert Langenbach
• 金额: $ 38.03万
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 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/7734559
• 关键词: Breeding; Cells; Chimera organism; DNA; Gastric ulcer; Genes; Genomics; Human; Infection; Inflammation; Knockout Mice; Lung; Lung Inflammation; Molecular; Mucins; Mus; Neomycin; PTGS2 gene; Pathway interactions; Pharmacology; Physiological; Polymerase Chain Reaction; Predisposition; Reporting; Role; Stomach; Ulcer; embryonic stem cell; homologous recombination; insight; malignant stomach neoplasm

A targeting construct containing the neomycin and HSTK genes and 10 kb of DNA genomic homology was constructed to disrupt the Muc5ac gene by homologous recombination. The construct was electroporated into mouse ES cells and 2 PCR positive clones of 170 clones analyzed were obtained. PCR and genomic Southern analysis verified targeting. Chimeras have been generated and 21 germline agouties have been obtained. The heterozygous Muc5ac mice are now being breed to generate wild type and Muc5ac-/- mice generated for lung inflammation and gastric ulceration studies.

构建了含有新霉素和HSTK基因以及10 kb的DNA基因组同源性的靶向构建体，以通过同源重组来破坏MUC5AC基因。将构建体电穿孔到小鼠ES细胞中，并获得了分析的170个克隆的2个PCR阳性克隆。 PCR和基因组南分析验证了靶向。已经产生了嵌合体，并获得了21种种系高素质。现在，杂合MUC5AC小鼠正在繁殖，以产生用于肺部炎症和胃溃疡研究的MUC5AC - / - 小鼠。