Purification and DNA Synthesis in Cell-Free Extracts: Properties of DNA Polymerase II
نویسندگان
چکیده
منابع مشابه
Purification and DNA synthesis in cell-free extracts: properties of DNA polymerase II.
The major DNA-synthesizing enzyme present in Pol A(1) (-)Escherichia coli (DNA polymerase II) has been purified to homogeneity as judged by polyacrylamide gel electrophoresis. The enzyme requires all four deoxynucleoside triphosphates, Mg(++), NH(4) (+), and native DNA for maximal activity. The enzyme activity is sensitive to sulfhydryl reagents and is insensitive to anti-DNA polymerase I antis...
متن کاملDNA synthesis in cell-free extracts of a DNA polymerase-defective mutant.
DNA SYNTHESIS IN CELL-FREE EXTRACTS OF A DNA POLYMERASE-DEFECTI VE MUTANT Thomas Kornberg and Malcolm L. Gefter Department of Biological Sciences Columbia University New York, New York 10027 Received August 4, 1970 Summary. A DNA-synthesizing activity has been isolated from an E. mmutant defective in DNA polvmerase. Like DNA polvmerase. the svstem requires the presence of all fbur’deoxynucleosi...
متن کاملInaccurate DNA Synthesis in Cell Extracts of Yeast Producing Active Human DNA Polymerase Iota
Mammalian Pol ι has an unusual combination of properties: it is stimulated by Mn(2+) ions, can bypass some DNA lesions and misincorporates "G" opposite template "T" more frequently than incorporates the correct "A." We recently proposed a method of detection of Pol ι activity in animal cell extracts, based on primer extension opposite the template T with a high concentration of only two nucleot...
متن کاملDeoxyribonucleic acid synthesis in cell-free extracts. IV. Purification and catalytic properties of deoxyribonucleic acid polymerase III.
DNA polymerase III, an enzyme essential for DNA replication, has been purified more than lO,OOO-fold from cellfree extracts of Escherichia coli. The enzyme, judged to be 10 to 20% pure, requires all four deoxynucleoside 5’-triphosphates, Mg++, ethanol, and native DNA for maximal activity. It is present in amounts sufficient to account for the in vivo rate of replication. DNA polymerase III acti...
متن کاملPurification and properties of DNA polymerase from Bacillus caldotenax.
A thermostable DNA polymerase was prepared from Bacillus caldotenax by using a four-step chromatography procedure. The protein exists as a monomer of M(r) 94,000, has a pI of 4.9 and has no associated 3'-5' or 5'-3'-exonuclease activities or endonuclease activity. The temperature optimum of the enzyme was about 70 degrees C and the pH for maximum activity was about 7.5. The enzyme has an absolu...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Proceedings of the National Academy of Sciences
سال: 1971
ISSN: 0027-8424,1091-6490
DOI: 10.1073/pnas.68.4.761