NR AMWQ
AU Yoshimoto,J.; Iinuma,T.; Ishiguro,N.; Horiuchi,M.; Imamura,M.; Shinagawa,M.
TI Comparative sequence analysis and expression of bovine PrP gene in mouse L-929 cells
QU Virus Genes 1992 Nov; 6(4): 343-56
PT journal article
AB A cDNA clone encoding bovine scrapie-associated fibril protein, PrP, from a bovine brain cDNA library and six amplified genomic DNA clones of bovine PrP were characterized. These clones possessed specific characteristics observed in other animal PrP genes. However, the bovine PrP was divided into two types by the number of repeats. One possessed four octapeptide repetitive sequences, like other animal PrP genes, and consisted of 256 amino acids; the other had five such repetitive sequences and 264 amino acids. The amino acid sequence of the former bovine PrP agreed with that of sheep PrP up to the 165th amino acid from the N-terminus. Bovine PrP cDNA introduced into mouse L-929 cells were stably expressed. The expression level of recombinant bovine PrP in the cells judged by immunofluorescence was higher than that of authentic mouse PrP. The recombinant PrP comigrated with authentic bovine PrP in SDS-polyacrylamide gel electrophoresis, suggesting that the recombinant product was fully glycosylated in L-929 cells. Distinct bundles of the intermediate filaments were frequently seen at the perinuclear region of the cells.
IN Eine Prion-cDNA aus einer Rindergehirn-cDNA-Bibliothek und 6 genomische Klone von amplifizierter Rinder-DNA wurden sequenziert. Sie kodierten zum Teil Prionproteine mit 4 Oktapeptidwiederholungen und 256 Aminosäuren, zum Teil aber auch 5 Oktapeptidwiederholungen und 264 Aminosäuren. Bis zur 165. Aminosäure stimmten die Rindersequenzen exakt mit den bei Schafen gefundenen überein. Die cDNA wurde in Mauszellen nicht nur exprimiert, sondern anscheinend auch genau wie in Rindern gylkosyliert.
MH Amino Acid Sequence; Animal; Base Sequence; Cattle; Cloning, Molecular; Female; Gene Expression; L Cells (Cell Line); Mice; Molecular Sequence Data; PrPsc Proteins; Prions/biosynthesis/chemistry/*genetics; Recombinant Proteins/biosynthesis; Sequence Analysis; Support, Non-U.S. Gov't; Transfection/genetics
AD Department of Veterinary Public Health, School of Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine, Japan.
SP englisch
PO USA