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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.

Additional information

Authors: CLERICI L, JRC Ispra (IT);SACCO M G, ITBA, CNR, Milano (IT);LUCCHINI F, ITBA, CNR, Milano (IT);FRANCESCHI C, Università di Modena (IT);FERRARI M, Università di Modena (IT);SCOVASSI A I, Istituto di Genetica Biochimica Evoluzionistica, CNR, Pavia (IT);NEGRONI M, Istituto di Genetica Biochimica Evoluzionistica, CNR, Pavia (IT);BERTAZZONI U, Istituto di Genetica Biochimica Evoluzionistica, CNR, Pavia (IT)
Bibliographic Reference: Paper presented: Fourth European Meeting on ADP-Ribosylation of Proteins, Pavia (IT), April 20-23, 1989 Text not available
Availability: Available from (1) as Paper EN 34753 ORA
Record Number: 199010011 / Last updated on: 1994-12-01
Original language: en
Available languages: en
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