Rapid isolation and characterization of 118 novel C2H2-type zinc finger cDNAs expressed in human brain |
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| Authors: | Becker Kavin G; Nagle James W; Canning Rachel D; Biddison William E; Ozato Keiko; Drew Paul D |
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| Institution: | 1Neuroimmunology Branch, National Institutes of Health Bethesda MD 20892, USA
2Section on Neurogenetics, National Institute of Neurological Diseases and Stroke, National Institutes of Health Bethesda MD 20892, USA
3Laboratory of Molecular Growth Regulation, National Institute of Child Health and Human Development, National Institutes of Health Bethesda MD 20892, USA |
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| Abstract: | C2H2-type zinc finger genes comprise one of the largest genefamilies in the human genome. These proteins are involved ingenetic regulation and development and are quite conserved throughoutevolution. The finger domains commonly contain the small linkerpeptide TGEKP between some finger units. Here, we report theisolation of 133 human zinc finger cDNAs, of which 118 are novel.These clones were isolated from human brain cDNA libraries usingoligonucleotide hybridization followed by expressed sequencetag (EST) analysis, sequencing from the conserved linker regionusing degenerate oligonucleotide primers. This directed partialsequencing approach to cDNA isolation and characterization,signature sequencing, combines the speed of EST automatic sequencingwith the focus of specific cDNA family analysis. Signature sequencingminimizes the generation of less informative random EST sequencesand provides a unique relative position for sequence comparison.We also show that there is an even distribution of these RNA5from this brain cDNA library, and that these cDNAs contain N-terminaldomains found in other zinc finger genes. This rapid focusedsequencing approach should be applicable to any family of cDNAscontaining short conserved signature peptide sequences. |
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