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.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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3T3 Cells
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Animals
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Base Sequence
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Chloramphenicol O-Acetyltransferase / genetics
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Chloramphenicol O-Acetyltransferase / metabolism
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Cloning, Molecular
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Exons
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Genes*
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Genomic Library
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Glucosylceramidase / genetics*
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Glucosylceramidase / metabolism
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Humans
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Kanamycin Kinase
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Mice
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Mice, Inbred BALB C
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Molecular Sequence Data
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Open Reading Frames
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Phosphotransferases / genetics
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Phosphotransferases / metabolism
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Promoter Regions, Genetic*
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Recombinant Fusion Proteins
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Restriction Mapping
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Sequence Homology, Nucleic Acid
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Transfection
Substances
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Recombinant Fusion Proteins
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Chloramphenicol O-Acetyltransferase
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Phosphotransferases
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Kanamycin Kinase
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Glucosylceramidase
Associated data
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GENBANK/M89944
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GENBANK/M89949
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GENBANK/M97581
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GENBANK/M97582
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GENBANK/M97583
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GENBANK/M97584
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GENBANK/M97585
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GENBANK/M97586
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GENBANK/M97587
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GENBANK/M97588
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GENBANK/M97589