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Studies on DNA replication in vitro by DNA polymerase-. alpha. /primase complex from embryonic chicken brain

Thesis/Dissertation ·
OSTI ID:7147073
DNA polymerase - {alpha} activity, characterized by its sensitivity to N-ethylmaleimide, high sedimentation coefficient, and acidic isoelectric point was found to decline with increasing embryonic age. Primase activity, the enzyme responsible for the initiation of DNA synthesis, was found to co-sediment with DNA polymerase - {alpha} activity on a continuous glycerol velocity gradient. Of all the single-stranded DNA templates tested, primase activity was found to be maximally active with poly dC. In the presence of ({alpha}-{sup 32}P) GTP, primase activity was found to catalyze the formation of an alkali-labile oligoriboguanylate, 15-20 nucleotides long. The chain length of the products were not altered by the presence or absence of dGTP. Primase activity was not inhibited by a high concentration of {alpha}- amanitin. Consistent with the finding of a decline of the activity level of DNA polymerase - {alpha}, the level of DNA polymerase - {alpha} antigen was also found to decrease with embryonic age, as evidenced by ELISA with a non-neutralizing monoclonal antibody, SJK 237-71.
Research Organization:
Notre Dame Univ., IN (USA)
OSTI ID:
7147073
Country of Publication:
United States
Language:
English

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