ADP-RIBOSYLATED PROTEINS IN MOUSE FERTILIZED EGGS MICROINJECTED WITH 32P-NAD
Fertilised eggs obtained from 3-4 weeks-old superovulated CD-1 mice were recovered at 1 cell stage by flushing the oviducts of the pregnant females the morning after the mating. 250-300 embryos were individually microinjected with about 10 pl of a solution containing 500-1000 pCi of 32P-NAD or 32P-NAD together with chemically synthesised pentadeca-oligodeoxinucleotides with sense or antisense sequences to cDNA of human poly(ADP-ribose) polymerase. The microinjected embryos were cultured overnight (18-22 hours, 5% CO(2), 37 C, 90% humidity), recovered in a small amount of PBS, subjected to 3 cycles of freezing and thawing and finally analysed on SDS-PAGE. The autoradiography of the gels showed that proteins with sizes of about 175, 110, 85, 60 and 15-25 KDa were 32P-NAD. When the oligomer with the sense sequence was added, on the contrary, the presence in the injected 32P-NAD solution of the antisense oligonucleotide appears to reduce significantly the labelling of the same proteins.
Bibliographic Reference: PAPER PRESENTED: FOURTH EUROPEAN MEETING ON ADP-RIBOSYLATION OF PROTEINS, PAVIA (IT), APRIL 20-23, 1989 TEXT NOT AVAILABLE
Record Number: 1989128003400 / Last updated on: 1990-11-09
Original language: EN
Available languages: en