A single P1 clone bearing three genes from human chromosome 11p15.5: HRC1, HRAS1, and RNH

Molecular genetic alterations of chromosome 11p15.5 are a common finding in human cancer. We previously reported the characterization of two cosmids representing a 55-kilobase (kb) region of DNA surrounding the protooncogene HRAS1. A cluster of genes was identified adjacent to this locus, and one of these genes, HRC1, was divergently transcribed 30 kb upstream from HRAS1. A recent report placed the gene for placental ribonuclease inhibitor (RNH, ribonuclease-angiogenin inhibitor) within 90 kb of HRAS1 by pulsed-field gel electrophoresis (PFGE) mapping. We used recombinant P1 bacteriophage clones for physical mapping to determine the position of RNH relative to the HRAS1 transcription unit and HRC1 on chromosome 11p15.5. PFGE and Southern analysis of genomic DNA suggested the order of the genes (HRC1-HRAS1-RNH). P1 clones confirmed this assignment, and placed RNH within 30-50 kb of the 3' end of HRAS1. Furthermore, a single 80-kb P1 clone that bears all three genes was isolated and clarified the Not I restriction map for the HRAS1-RNH interval. Their close physical association was predicted by simple screening of an arrayed P1 library; the clone containing all three genes was selected from multiple positive signals obtained for each HRC1 and RNH because it mapped to the same library-well address.