Chang J G, Scarpa A, Eddy R L, Byers M G, Harris A S, Morrow J S, Watkins P, Shows T B, Forget B G
Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06510.
Genomics. 1993 Aug;17(2):287-93. doi: 10.1006/geno.1993.1323.
A 96-bp synthetic oligonucleotide corresponding to an amino acid sequence near the N-terminus of erythroid beta-spectrin was used to screen a human genomic library, and two overlapping recombinants were isolated. DNA sequence analysis established that the genomic fragment encoded beta-fodrin, the nonerythroid form of beta-spectrin, by correlation to a known amino acid sequence of human brain beta-fodrin. The genomic DNA contained regions that cross-hybridized with an erythroid beta-spectrin cDNA probe, and the DNA sequence of these regions revealed a high degree of identity with that of erythroid beta-spectrin and a similar exon/intron organization. A single-copy DNA fragment of the beta-fodrin genomic clone was used to screen a lymphoid cell cDNA library and two recombinants were isolated. The composite DNA sequence of these various genomic and cDNA clones encoded almost all of the first twelve 106 amino acid repeat segments of beta-fodrin that shared 58% identity and 75.5% similarity with the amino acid sequence of beta-spectrin and 66% identity with the nucleotide sequence of beta-spectrin cDNA. The chromosomal localization of the gene was determined to be chromosome 2 by hybridization of a single-copy probe derived from the cloned genomic DNA to DNA of a panel of somatic hybrid cell lines, and in situ hybridization localized the gene to band 2p21. beta-Fodrin was assigned the gene symbol SPTBN1.
使用一段与红系β-血影蛋白N端附近氨基酸序列相对应的96个碱基对的合成寡核苷酸筛选人基因组文库，分离出两个重叠的重组体。DNA序列分析表明，通过与已知的人脑β-肌动蛋白氨基酸序列相关联，该基因组片段编码β-肌动蛋白，即β-血影蛋白的非红系形式。基因组DNA包含与红系β-血影蛋白cDNA探针交叉杂交的区域，这些区域的DNA序列与红系β-血影蛋白的序列高度同源，并且具有相似的外显子/内含子组织。β-肌动蛋白基因组克隆的单拷贝DNA片段用于筛选淋巴细胞cDNA文库，分离出两个重组体。这些不同的基因组和cDNA克隆的复合DNA序列编码了β-肌动蛋白几乎所有的前十二个106个氨基酸的重复片段，这些片段与β-血影蛋白的氨基酸序列具有58%的同源性和75.5%的相似性，与β-血影蛋白cDNA的核苷酸序列具有66%的同源性。通过将源自克隆基因组DNA的单拷贝探针与一组体细胞杂种细胞系的DNA杂交，确定该基因的染色体定位为2号染色体，原位杂交将该基因定位到2p2 l带。β-肌动蛋白被赋予基因符号SPTBN1。
