Isolation and expression of a rat liver cDNA encoding phosphoglucomutase.
A cDNA encoding phosphoglucomutase (PGM) has been isolated from a rat liver cDNA library following screening with a polymerase chain reaction product. The cDNA was found to contain a 53-base-pair (bp) 5' untranslated region (5' UTR), a single start codon and consensus initiation sequence, an open reading frame (ORF) of 1686 bp, and a 3' untranslated tail. A comparison to the rabbit and human muscle PGM cDNAs [Whitehouse et al., Proc. Natl. Acad. Sci. USA 89 (1992) 411-415] showed 90% identity of rat cDNA to both, while a comparison to the deduced amino acid sequences showed 97 and 96% identity, respectively. Northern blot analyses determined that PGM was encoded by a single mRNA in rat liver, of approximately 2.2 kb. Following transfection of COS-7 cells with a plasmid containing the entire PGM ORF, indirect immunofluorescence analyses using a PGM-specific monoclonal antibody determined that approximately 5% of the cells displayed 50-100 times greater fluorescence than that seen in the remainder of the cells or in mock transfects. The enhanced production of PGM was also demonstrated by Western blotting and by enzymatic activity assays.
Rivera
A A
AA
Department of Cell Biology, University of Alabama at Birmingham 35294-0005.
Elton
T S
TS
Dey
N B
NB
Bounelis
P
P
Marchase
R B
RB
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA; D005455:Fluorescent Antibody Technique; D006801:Humans; D007438:Introns; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010733:Phosphoglucomutase / Q000096:biosynthesis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D051381:Rats
D004247:DNA; D010733:Phosphoglucomutase