Tse W T, Menninger J C, Yang-Feng T L, Francke U, Sahr K E, Lux S E, Ward D C, Forget B G
Department of Human Genetics, Yale University School of Medicine, New Haven, Connecticut 06510.
Genomics. 1991 Aug;10(4):858-66. doi: 10.1016/0888-7543(91)90173-c.
Immunoreactive isoforms of erythrocyte ankyrin have been shown to be present in a variety of nonerythroid tissues. Isolation of the genes that encode these isoforms will clarify their relationship to erythrocyte ankyrin. Using an erythrocyte ankyrin cDNA clone as a hybridization probe, we screened a human genomic library and isolated a clone that hybridizes with the probe at low stringency but not at high stringency. Partial nucleotide sequence of the clone revealed the presence of a 99-bp segment that is homologous to an exon of the erythrocyte ankyrin gene. Northern analysis showed that a labeled fragment of the clone hybridized to a 7-kb message in RNA of fetal brain but not of erythroid cells, suggesting that this clone is part of a novel gene that is expressed predominantly in nonerythroid tissue. Comparison of the sequence of the genomic clone with that of a recently isolated cDNA clone for brain ankyrin (Otto et al., 1989) showed identity of 96 of 99 bp between the putative exon and a segment of the cDNA clone (V. Bennett, personal communication, 1991), suggesting that the genomic clone is part of a gene for nonerythroid ankyrin, which we have designated ANK2. By analysis of somatic cell hybrids and fluorescence in situ hybridization, we assigned ANK2 to human chromosome 4 at a position equivalent to bands 4q25-q27.
红细胞锚蛋白的免疫反应性同工型已被证明存在于多种非红细胞组织中。编码这些同工型的基因的分离将阐明它们与红细胞锚蛋白的关系。我们使用红细胞锚蛋白cDNA克隆作为杂交探针,筛选了一个人类基因组文库,并分离出一个在低严谨度下与探针杂交但在高严谨度下不杂交的克隆。该克隆的部分核苷酸序列显示存在一个与红细胞锚蛋白基因外显子同源的99bp片段。Northern分析表明,该克隆的一个标记片段与胎儿脑RNA中的一个7kb信息杂交,但与红细胞系细胞的RNA不杂交,这表明该克隆是一个主要在非红细胞组织中表达的新基因的一部分。将基因组克隆的序列与最近分离的脑锚蛋白cDNA克隆(Otto等人,1989)的序列进行比较,发现在假定的外显子和cDNA克隆的一个片段之间99bp中有96bp相同(V. Bennett,个人交流,1991),这表明该基因组克隆是非红细胞锚蛋白基因的一部分,我们将其命名为ANK2。通过对体细胞杂种的分析和荧光原位杂交,我们将ANK2定位到人类4号染色体上相当于4q25 - q27带的位置。