Abstract
The crystallins are highly conserved structural proteins universally found in the eye lens of all vertebrate species. In mammals, three immunologically distinct classes are present, alpha-, beta-, and gamma-crystallins, and each class represents a multigene family. The alpha-crystallin gene family consists of alpha 1-crystallin (CRYA1) and alpha 2-crystallin (CRYA2) genes (previously designated alpha A- and alpha B-crystallin, respectively), which show extensive sequence homology. We constructed a synthetic oligonucleotide probe of 25 bases corresponding to a specific region of the human alpha 1-crystallin gene sequence. This 25-mer probe bears little sequence homology to human alpha 2-crystallin gene and does not cross-hybridize to alpha 2-crystallin sequences in Southern blot analysis. Using this unique synthetic probe, we have demonstrated the identity of the alpha 1-crystallin gene in human genomic DNA. In addition, we have also confirmed its chromosomal location on human chromosome 21. Finally, we have regionally localized the gene to q22.3 by using both Southern blot analysis of a panel of cell hybrids containing different parts of human chromosome 21, and in situ hybridization to metaphase chromosomes. The use of synthetic oligonucleotide probes specific for individual genes should be useful in identifying and mapping members of multigene families.
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