Bloom M L, Kaysser T M, Birkenmeier C S, Barker J E
Jackson Laboratory, Bar Harbor, ME 04609.
Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):10099-103. doi: 10.1073/pnas.91.21.10099.
The jaundiced, ja/ja, mouse mutant has a severe hemolytic anemia associated with a deficiency of beta-spectrin in erythrocyte ghosts. Genes for the disease phenotype and beta-spectrin colocalize on Chromosome 12. beta-Spectrin mRNA is not detected in reticulocytes or in brain from newborn mutant mice. To locate the nucleotide sequence alteration, the erythroid beta-spectrin transcript from mutant spleen was amplified by reverse transcription PCR and sequenced. A C-to-T alteration is present in the mutant transcript and produces a premature stop codon from an arginine codon in mRNA encoding repeat 9 of beta-spectrin at amino acid position 1160. The point mutation introduces a Dde I site that is present in PCR-amplified DNA of ja/ja and ja/+ mice but not of +/+ control mice from the strain of origin, 129/Sv, or from the two strains, WB/Re and C57BL/6J, in which the mutation has been fixed by over 53 generations of backcrossing. The genetic data confirm that the point mutation is responsible for the severe reductions in beta-spectrin mRNA of jaundiced mice.
黄疸型(ja/ja)小鼠突变体患有严重的溶血性贫血,与红细胞膜中β-血影蛋白缺乏有关。该疾病表型基因和β-血影蛋白基因共定位于第12号染色体上。在新生突变小鼠的网织红细胞或脑中未检测到β-血影蛋白mRNA。为了定位核苷酸序列改变,通过逆转录PCR扩增突变体脾脏中的红系β-血影蛋白转录本并进行测序。突变转录本中存在一个C到T的改变,在编码β-血影蛋白重复序列9的mRNA中,从第1160位氨基酸的精氨酸密码子产生一个提前终止密码子。该点突变引入了一个Dde I位点,该位点存在于黄疸型(ja/ja)和杂合型(ja/+)小鼠的PCR扩增DNA中,但在来自原始品系129/Sv或两个品系WB/Re和C57BL/6J的野生型(+/+)对照小鼠的PCR扩增DNA中不存在,在这两个品系中该突变已经通过超过53代的回交而固定。遗传数据证实该点突变是黄疸型小鼠β-血影蛋白mRNA严重减少的原因。