Characterization of the mammalian DNA polymerase gene and protein. Annual progress report
We have purified and characterized at least three DNA polymerases from Chinese hamster ovary (CHO) cell line Kl in order to evaluate the roles of different polymerases in eukaryotic DNA replication. Pol {alpha} was the most abundant among different polymerase activities and it was neutralized by a monoclonal antibody raised against human pol {alpha}. Pol {var_epsilon} was separated from pol {alpha} and pol {delta} activities using DEAE Sephacell, phosphocellulose and hydroxylapatite columns. The enzyme proved to be sensitive to aphidicolin and carbonyldiphosphonate and was not stimulated by PCNA- Pol {delta} was the least abundant among the three enzymes. It was sensitive to aphidicolin and carbonyidiphosphonate and was stimulated by PCNA. it had a preference for template/primer poly (dA-dT). Based on DNA sequence data of different eukaryotic polymerases PCR primers complementary to two neighboring synthesized. In PCR experiments several products were obtained which are assumed to be specific for the CHO polymerases. We have also analyzed a large number of aphidicolin resistant mutants of CHO to identify mutants with altered DNA polymerases.
- Research Organization:
- South Carolina Univ., Columbia, SC (United States)
- Sponsoring Organization:
- USDOE, Washington, DC (United States)
- DOE Contract Number:
- FG09-90ER61040
- OSTI ID:
- 10165626
- Report Number(s):
- DOE/ER/61040-2; ON: DE93017270
- Resource Relation:
- Other Information: PBD: Jan 1992
- Country of Publication:
- United States
- Language:
- English
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Characterization of the mammalian DNA polymerase gene and protein. Annual progress report
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