IDENTIFICATION OF ZINC-FINGER MESSENGER-RNAS USING DOMAIN-SPECIFIC DIFFERENTIAL DISPLAY

An oligonucleotide primer specific for a conserved amino acid region o f the Cys(2)/His(2) zinc finger proteins was used in conjunction with mRNA differential display to amplify related mRNAs from a human ovaria n cancer cell Line. Six of the 12 cDNAs analyzed from the differential display polyacrylamide gel exhibited zinc finger homology at the nucl eotide and predicted amino acid sequence level. None of these cDNA fra gments, however, shared complete homology with genes encoding any know n zinc finger proteins. All 6 cDNA fragments with zinc finger homology had a poly-A tail and 3 of these fragments contained a putative polya denylation signal. Northern blot analysis was performed using radiolab eled probes prepared from the 12 cDNA fragments. Two Of the 6 cDNA fra gments with zinc finger homology hybridized to 3.6- and 6.0-kb mRNAs. In addition, 2 of the fragments which did not contain significant homo logy to zinc finger or any other known sequences hybridized to 4.1- an d 5.8-kb mRNAs. These results suggest that domain-specific differentia l display may be a useful approach for the identification of novel gen e family members as well as for the analysis of changes in gene expres sion of family members between related cell Lines or tissue samples. ( C) 1996 Academic Press, Inc.