Cloning a cDNA for the lysosomal alpha-glucosidase.
Messenger RNA was isolated from monkey testes and size-fractionated on sucrose gradients. In vitro translation of these mRNA fractions resulted in nascent, labeled alpha-glucosidase that could be precipitated with anti human alpha-glucosidase antiserum. A cDNA library was constructed from the most enriched fraction. The library was screened with cDNA made from mRNA obtained from immunoselected polysomes. Five cross-hybridizing clones were isolated and identified by their selection of alpha-glucosidase mRNA, as shown by hybrid released translation and further by their ability to hybridize with DNA from human chromosome 17, on which the gene coding for acid alpha-glucosidase is located.
D000818:Animals; D002902:Chromosomes, Human, 16-18; D003001:Cloning, Molecular; D004247:DNA / Q000096:biosynthesis*; D004274:DNA, Recombinant / Q000096:biosynthesis; D005959:Glucosidases / Q000235:genetics*; D006801:Humans; D007163:Immunosorbent Techniques; D008247:Lysosomes / Q000201:enzymology*; D008251:Macaca; D008297:Male; D051379:Mice; D009693:Nucleic Acid Hybridization; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D013737:Testis / Q000032:analysis; D000520:alpha-Glucosidases / Q000235:genetics*
D004274:DNA, Recombinant; D012333:RNA, Messenger; D004247:DNA; D005959:Glucosidases; D000520:alpha-Glucosidases