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Cloning, Expression, and Mapping of Ribonucleases H of Human and Mouse Related to Bacterial RNase HI

1998; Elsevier BV; Volume: 53; Issue: 3 Linguagem: Inglês

10.1006/geno.1998.5497

1089-8646

Susana M. Cerritelli, Robert J. Crouch,

RNA Interference and Gene Delivery

We identified two human sequences and one mouse sequence in the database of expressed sequence tags that are highly homologous to the N-terminal sequence of eukaryotic RNases H1. The cDNAs for humanRNASEH1and mouseRnaseh1were obtained, their nucleotide sequences determined, and the proteins expressed inEscherichia coliand partially purified. Both proteins have RNase H activityin vitroand they bind to dsRNA and RNA–DNA hybrids through the N-terminal conserved motif present in eukaryotic RNases H1. TheRNASEH1gene is expressed in all human tissues at similar levels, indicating that RNase H1 may be a housekeeping protein. The humanRNASEH1and mouseRnaseh1cDNAs were used to isolate BAC genomic clones that were used as probes for fluorescencein situhybridization. The human gene was localized to chromosome 17p11.2 and the mouse gene to a nonsyntenic region on chromosome 12A3. The chromosomal location and possible disease association of the humanRNASEH1gene are discussed.