ISOLATION AND CHARACTERIZATION OF TRANSCRIBED SEQUENCES FROM A CHROMOSOME-16 HN-CDNA LIBRARY AND THE PHYSICAL MAPPING OF GENES AND TRANSCRIBED SEQUENCES USING A HIGH-RESOLUTION SOMATIC-CELL PANEL OF HUMAN CHROMOSOME-16

A hn-cDNA (heteronuclear complementary DNA) library was constructed fr om a mouse/human somatic cell hybrid, CY18, which contains chromosome 16 as the only human chromosome. Hexamer primers constructed from cons ensus 5' intron splice sequences were used to generate cDNA from the i mmature unspliced mRNA. The resulting cDNA library was screened with a total human DNA probe to identify potential human clones. Rescreening was necessary, and use of a mouse-derived clone with homology to 7SL RNA proved successful in eliminating the majority of mouse clones. Thi rteen clones had open reading frames, and of those, five showed homolo gy to human sequences in Gen-Bank. Two clones had homology to random p artially sequenced cDNAs, one clone was likely to be a GRP78 pseudogen e, one clone mapped the PHKG2 gene to 16p11.2-16p12.1, and one clone h ad homology to human S13 ribosomal protein. All clones except the latt er were mapped to a high-resolution somatic cell panel. Although isola tion of human chromosome 16 genes from this library was successful, it was apparent that cDNA synthesis was initiated at sites other than in tron splice sites, presumably by mispairing of the hexamers. (C) 1994 Academic Press, Inc.