Document Type
Article
Publication Date
11-25-1990
Department
Chemistry and Biochemistry
School
Mathematics and Natural Sciences
Abstract
A DNA polymerase has been highly purified from Anacystis nidulans R2. Electrophoretic analysis in sodium dodecyl sulfate-polyacrylamide gels revealed that the final fraction contains three bands of Mr 107,000, 93,000, and 51,000, respectively. Analysis of purified DNA polymerase activity in situ indicates that of the three polypeptides the Mr 107,000 species has the catalytic activities. The native molecular weight of the enzyme was estimated by glycerol gradient sedimentation to be 100,000. The enzyme has an absolute requirement for a divalent cation. Mg2+ can be replaced with Mn2+, but the DNA polymerase is less active. Potassium chloride stimulates the enzyme, while potassium phosphate has no apparent effect. The enzyme is active over a pH range from 7.5 to 9.5 in 50mM Tris-HCI buffer. The ability of the cyanobacterial DNA polymerase to use activated DNA as a template, Its associated 3′ →5′ and 5′→3′ exonuclease activities, as well as its resistance to N-ethylmaleimide, dideoxynucleotides, arabinosyl-CTP and aphidicolln suggest a similarity between this enzyme and E. coli DNA polymerase I. This is the first characterization of a DNA polymerase from a cyanobacterium.
Publication Title
Nucleic Acids Research
Volume
18
Issue
22
First Page
6659
Last Page
6663
Recommended Citation
Lin, H.,
Cannon, G. C.,
Heinhorst, S.
(1990). Purification and Characterization of a DNA Polymerase From the Cyanobacterium Anacystis nidulans R2. Nucleic Acids Research, 18(22), 6659-6663.
Available at: https://aquila.usm.edu/fac_pubs/7503
Comments
Originally published in Nucleic Acids Research, Vol. 18, No. 22