Birkenmeier C S, Bodine D M, Repasky E A, Helfman D M, Hughes S H, Barker J E
Proc Natl Acad Sci U S A. 1985 Sep;82(17):5671-5. doi: 10.1073/pnas.82.17.5671.
A cDNA clone for nonerythroid alpha-spectrin was identified by direct immunological screening of a chicken smooth muscle cDNA library. A library prepared in the expression plasmids pUC8 and pUC9 was screened with an antiserum specific for chicken alpha-spectrin. Blots of poly(A)+ RNA from various tissues of chicken and mouse show that the cDNA hybridizes to an 8-kilobase mRNA. The cDNA hybridizes to a single-copy sequence on Southern blots of chicken genomic DNA. The complete nucleic acid sequence of the clone has a single 1419-base open reading frame. The derived amino acid sequence is organized into two partial and three complete 106-amino-acid repeats that show homology to the repeats described for human erythroid alpha- and beta-spectrin. Immunological and biochemical data indicate that chicken nonerythroid and human erythroid alpha-spectrin are two of the more widely diverged members of the spectrin family of proteins. In this respect, the degree of homology found between them was unexpected. Our data suggest a common evolutionary origin for these two alpha-spectrins and allow some predictions concerning spectrin gene structure.
通过对鸡平滑肌cDNA文库进行直接免疫筛选,鉴定出了非红细胞α-血影蛋白的一个cDNA克隆。用对鸡α-血影蛋白具有特异性的抗血清筛选在表达质粒pUC8和pUC9中构建的文库。来自鸡和小鼠各种组织的聚腺苷酸加尾RNA的印迹显示,该cDNA与一个8千碱基的mRNA杂交。该cDNA与鸡基因组DNA的Southern印迹上的单拷贝序列杂交。该克隆的完整核酸序列有一个1419个碱基的单一开放阅读框。推导的氨基酸序列被组织成两个部分重复和三个完整的106个氨基酸的重复,这些重复与人类红细胞α-和β-血影蛋白中描述的重复序列具有同源性。免疫学和生化数据表明,鸡非红细胞α-血影蛋白和人类红细胞α-血影蛋白是血影蛋白家族中分化程度较高的两个成员。在这方面,它们之间发现的同源程度是出乎意料的。我们的数据表明这两种α-血影蛋白有共同的进化起源,并对血影蛋白基因结构做出了一些预测。