Molecular cloning of an alpha-glucosidase-like gene from Penicillium minioluteum and structure prediction of its gene product.
The dexC cDNA, which is expressed in dextran-containing medium by the filamentous fungus Penicillium minioluteum, was cloned and sequence characterized. The cDNA sequence comprises 1859 bp plus a poly (A) tail, coding for a predicted protein of 597 amino acids. The genomic counterpart was isolated by PCR, finding three introns in its sequence. The dexC gene was located by Southern blot in the same 9-kb fragment that the previously isolated dextranase-encoding gene (dexA). Sequence analysis revealed that the deduced DexC protein belongs to glycosyl hydrolase family 13, showing a high sequence identity (58%) with Aspergillus parasiticus alpha-1,6-glucosidase. In addition, the high sequence identity (51%) between DexC protein and oligo-1,6-glucosidase of Bacillus cereus, with three-dimensional (3D) structure determined, leads us to proposed a 3D model for the structural core of DexC protein.
García
B
B
Centro de Ingeniería Genética y Biotecnología (CIGB), Havana, Cuba.
Castellanos
A
A
Menéndez
J
J
Pons
T
T
D000595:Amino Acid Sequence; D001230:Aspergillus / Q000201:enzymology; D001407:Bacillus / Q000201:enzymology; D001483:Base Sequence; D015139:Blotting, Southern; D020134:Catalytic Domain; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004247:DNA / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D016208:Databases, Factual; D003910:Dextranase / Q000378:metabolism; D005656:Fungal Proteins; D005959:Glucosidases / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000235:genetics*; D007438:Introns; D008957:Models, Genetic; D008958:Models, Molecular; D008969:Molecular Sequence Data; D005810:Multigene Family; D009834:Oligo-1,6-Glucosidase / Q000378:metabolism; D010407:Penicillium / Q000201:enzymology* / Q000235:genetics*; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D000520:alpha-Glucosidases / Q000737:chemistry* / Q000235:genetics*
D018076:DNA, Complementary; D005656:Fungal Proteins; D011994:Recombinant Proteins; D004247:DNA; C423005:DexC protein, Penicillium minioluteum; D005959:Glucosidases; D006026:Glycoside Hydrolases; D009834:Oligo-1,6-Glucosidase; D003910:Dextranase; D000520:alpha-Glucosidases; C023149:exo-1,6-alpha-glucosidase