MUTANT HEMOGLOBINS THAT ALLOW HBS TO SICKLE

将产生一群含有血红蛋白和氧气的小鼠 与人类相似的联想-解离特性 镰状细胞血红蛋白(HBS)。这将通过培育一种 在血红蛋白基因座上携带两个突变的小鼠。 Hbag2α-珠蛋白单倍型纯合的小鼠将 与Hbbs2β-珠蛋白纯合的小鼠交配 单倍型。将对F2后代进行筛查，以识别出 α-和β-珠蛋白基因突变的双重纯合。 Hbag2/Hbag2；Hbbs2/Hbbs2小鼠的血红蛋白将会有氧 缔合-解离性质类似于HBS。这个 老鼠将被称为MHOAH。MHOAH小鼠将被转基因用于 人类甲型和镰状细胞或镰状细胞安的列斯-贝塔- 利用珠蛋白基因制造镰状细胞转基因小鼠模型 疾病。转基因将被导入MHOAH小鼠体内 直接与携带转基因的小鼠种群繁殖 或者直接将微小的DNA构建物注射到 MHOAH小鼠受精卵的雄性原核。携带的老鼠 转基因将通过Southern blotting进行鉴定。转基因 表达高水平转基因的小鼠将被识别出来 通过血液溶血物的电泳法。氧的缔合- MHOAH小鼠体内血红蛋白的解离和凝胶化特性 表达高水平的HBS或HBS Antilles的人将被研究。 表达高水平HBS Antilles的转基因小鼠非常 很可能是镰状细胞病的动物模型。哈佛商学院安的列斯群岛 具有比HBS更低的溶解度和更低的氧亲和力。关于 60%的HBS安的列斯群岛将在40毫米汞下脱氧 氧分压；只有25%的高氧亲和力 MHOAH小鼠的血红蛋白在相同的氧气条件下脱氧 紧张。这些条件导致了HBA/S的镰状细胞病 Antilles杂合子，有可能产生HBs凝胶化 转基因MHOAH小鼠红细胞的倾斜和镰刀样改变。 镰状细胞性贫血的小鼠模型将有助于研究 论该病的病理生理学及发病机制 抗感冒药的检测。

A colony of mice will be produced that has hemoglobin with oxygen association-dissociation properties that are similar to human sickle cell hemoglobin (HbS). This will be done by breeding a stock of mice that carry two mutations at the hemoglobin loci. Mice that are homozygous for the Hbag2 alpha-globin haplotype will be mated with mice that are homozygous for the Hbbs2 beta-globin haplotype. F2 progeny will be screened to identify mice that are doubly homozygous for the alpha- and beta-globin gene mutations. The hemoglobin of Hbag2/Hbag2;Hbbs2/Hbbs2 mice will have oxygen association-dissociation properties similar to that of Hbs. The mice will be called MHOAH. MHOAH mice will be made transgenic for the human alpha- and sickle cell or sickle cell Antilles beta- globin genes to produce a transgenic mouse model for sickle cell disease. The transgenes will be introduced into MHOAH mice directly by breeding with stocks of mice that carry the transgenes or directly by microinjecting minilocus constructs of DNA into the male pronucleus of fertilized eggs of MHOAH mice. Mice that carry the transgenes will be identified by Southern blotting. Transgenic mice that express high levels of the transgenes will be identified by electrophoresis of blood hemolysates. The oxygen association- dissociation and gelation properties of hemoglobins in MHOAH mice that express high levels of HbS or HbS Antilles will be studied. Transgenic mice that express high levels of HbS Antilles are very likely to be an animal model for sickle cell disease. HbS Antilles has a lower solubility and a lower oxygen affinity than HbS. About 60 percent of the HbS Antilles will be deoxygenated at 40 mm of Hg oxygen tension; only 25 percent of the high oxygen affinity hemoglobin of MHOAH mouse would be deoxygenated at the same oxygen tension. These conditions produce sickle cell disease in HbA/S Antilles heterozygotes and are likely to produce gelation of HbS Antilles and sickling of erythrocytes in transgenic MHOAH mice. The mouse model for sickle cell anemia would facilitate research on the pathophysiology of the disease and on the development and testing of anti-sickling drugs.