title: Molecular and functional characterization of the murine glucocerebrosidase gene.
abstract: A genomic clone of glucocerebrosidase (D-glucosyl-N-acyl-sphingosine glucohydrolase; E.C. 3.2.1.45) purified from a genomic library derived from a Balb/c mouse was analyzed by restriction mapping and nucleotide sequencing of its promoter and protein coding regions. Promoter activity was functionally assessed by ligation of a 2 kb glucocerebrosidase fragment to the protein coding segment of a bacterial neomycin resistance gene. Smaller segments of the 5' flanking sequence were then analyzed for their ability to initiate transcription of the chloramphenicol acetyltransferase reporter gene. A 319 bp Eco RI-Bgl II fragment (containing 259 bp upstream of the cDNA 5' limit) ligated to the chloramphenicol acetyltransferase open reading frame produced considerable activity.
authors.affiliation: Developmental and Metabolic Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: E D
authors.initials: ED
authors.lastname: Carstea
authors.forename: G J
authors.initials: GJ
authors.lastname: Murray
authors.forename: R R
authors.initials: RR
authors.lastname: O'Neill
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D005091:Exons; D005796:Genes; D015698:Genomic Library; D005962:Glucosylceramidase / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D019868:Kanamycin Kinase; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010770:Phosphotransferases / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection
chemical_list: D011993:Recombinant Fusion Proteins; D015500:Chloramphenicol O-Acetyltransferase; D010770:Phosphotransferases; D019868:Kanamycin Kinase; D005962:Glucosylceramidase