Wawrzyniak C J, Popp R A
Biochem Genet. 1986 Apr;24(3-4):259-72. doi: 10.1007/BF00502793.
Dot blot hybridization was used to determine the relative amounts of the beta-major and beta-minor globin RNAs present in reticulocytes of mice at 14.5 and 17.5 days of gestation, newborns, and adults of the Hbab/Hbab;Hbbs2/Hbbs2 globin genotype. RNAs isolated from embryonic yolk sac, fetal liver, and adult reticulocytes were hybridized with the following labeled DNA probes: alpha-1, beta-minor specific, and beta-major specific. The level of beta-sminor RNA in reticulocytes at 14.5 and 17.5 days of gestation is nearly the same as in induced reticulocytes of adult mice. In contrast, the level of beta-s2major RNA in reticulocytes at 14.5 days of gestation is 0.23 X and at 17.5 days of gestation is 0.66 X the amount found in reticulocytes of adult mice. These results correlate well with earlier observations that the beta-sminor globin gene approaches its normal adult level of expression by 14.5 days of gestation, whereas the beta-s2major globin gene expression increases between 14.5 days of gestation and 6 days postnatally. They indicate that the differential expression of beta-sminor and beta-s2major globins during development is regulated at the level of transcription. Expression of the beta-minor globin gene in reticulocytes of adult normal mice is not maximal, however, because the levels of the beta-minor globin and its RNA are increased further in reticulocytes of thalassemic mice.
斑点杂交法用于测定妊娠14.5天和17.5天的小鼠、新生小鼠以及Hbab/Hbab;Hbbs2/Hbbs2珠蛋白基因型成年小鼠网织红细胞中β-主要和β-次要珠蛋白RNA的相对含量。从胚胎卵黄囊、胎儿肝脏和成年网织红细胞中分离的RNA与以下标记的DNA探针杂交:α-1、β-次要特异性探针和β-主要特异性探针。妊娠14.5天和17.5天的网织红细胞中β-次要RNA的水平与成年小鼠诱导网织红细胞中的水平几乎相同。相比之下,妊娠14.5天时网织红细胞中β-主要RNA的水平是成年小鼠网织红细胞中该水平的0.23倍,妊娠17.5天时是0.66倍。这些结果与早期观察结果很好地相关,即β-次要珠蛋白基因在妊娠14.5天时接近其正常成年表达水平,而β-主要珠蛋白基因表达在妊娠14.5天至出生后6天之间增加。它们表明β-次要和β-主要珠蛋白在发育过程中的差异表达是在转录水平上调控的。然而,成年正常小鼠网织红细胞中β-次要珠蛋白基因的表达并非最大,因为地中海贫血小鼠网织红细胞中β-次要珠蛋白及其RNA的水平进一步增加。