title: Feature mapping of the HLA class I region: localization of the POU5F1 and TCF19 genes.
abstract: The class I region of the human leukocyte antigen (HLA) complex located on chromosome 6p21.3 is gene dense. To define the gene content of the class I region, we are constructing genomic DNA feature maps. Here we report mapping of the POU5F1 and TCF19 genes to an approximately 0.2-Mb region between the HLA-C and the S genes. Localization of these genes was facilitated by subcloning genomic DNA fragments from the 0.2-Mb region into a transposon gamma delta-based vector, selecting transposon-mediated deletions in vivo in Escherichia coli, and sequencing a nested subset chosen for their uniform distribution of deletion endpoints. The POU5F1 and TCF19 genes are located approximately 130 kb telomeric of the HLA-C locus, approximately 0.6 kb apart from each other. Complete sequencing of a 5.5-kb EcoRI fragment containing the TCF19 gene revealed that it is composed of three exons, bounded by consensus splice signals. These experiments illustrate that the transposon-based nested deletion sequencing method provides an easy and efficient approach to feature mapping genomic fragments and to high-resolution analysis of selected subportions.
authors.affiliation: Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
authors.forename: B R
authors.initials: BR
authors.lastname: Krishnan
authors.forename: I
authors.initials: I
authors.lastname: Jamry
authors.forename: D D
authors.initials: DD
authors.lastname: Chaplin
mesh_terms: D001483:Base Sequence; D002874:Chromosome Mapping / Q000379:methods*; D004268:DNA-Binding Proteins / Q000235:genetics*; D015395:Histocompatibility Antigens Class I / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D050814:Octamer Transcription Factor-3; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific) / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D004268:DNA-Binding Proteins; D015395:Histocompatibility Antigens Class I; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C098583:TCF19 protein, human; D014157:Transcription Factors; C020098:DNA modification methylase EcoRI; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific)