Switch to
Predicate | Object |
---|---|
rdf:type | |
lifeskim:mentions | |
pubmed:issue |
22
|
pubmed:dateCreated |
1987-9-17
|
pubmed:abstractText |
Error rates for conventionally purified DNA polymerase-alpha from calf thymus, chicken, and human sources have been reported to be one in 10,000 to one in 40,000 nucleotides incorporated. Isolation of polymerase-alpha by immunoaffinity chromatography yields a multiprotein high molecular weight replication complex that contains an associated DNA primase (Wong, S. W., Paborsky, L. R., Fisher, P. A., Wang, T. S-F., and Korn, D. (1986) J. Biol. Chem. 261, 7958-7968). We have isolated DNA polymerase-primase complexes from calf thymus, from a human lymphoblast cell line (TK-6), and from Chinese hamster lung cells (V-79) using two different methods of immunoaffinity chromatography. These enzyme complexes are 12- to 20-fold more accurate than conventionally purified calf thymus DNA polymerase-alpha when assayed using the phi X174am3 fidelity assay; estimated error rates are one in 460,000 to one in 830,000 nucleotides incorporated when the enzyme complex is freshly isolated. The polymerase-primase complex from calf thymus exhibited no detectable 3'----5' exonuclease activity using a heteroduplex substrate containing a single 3'-terminal mismatched nucleotide. Upon prolonged storage at -70 degrees C, the error rate of the immunoaffinity-purified calf thymus DNA polymerase-primase complex increases to about one in 50,000 nucleotides incorporated, an error rate similar to that exhibited by conventional isolates of DNA polymerase-alpha.
|
pubmed:grant | |
pubmed:language |
eng
|
pubmed:journal | |
pubmed:citationSubset |
IM
|
pubmed:chemical | |
pubmed:status |
MEDLINE
|
pubmed:month |
Aug
|
pubmed:issn |
0021-9258
|
pubmed:author | |
pubmed:issnType |
Print
|
pubmed:day |
5
|
pubmed:volume |
262
|
pubmed:owner |
NLM
|
pubmed:authorsComplete |
Y
|
pubmed:pagination |
10824-30
|
pubmed:dateRevised |
2007-11-14
|
pubmed:meshHeading |
pubmed-meshheading:3038898-Animals,
pubmed-meshheading:3038898-Bacteriophage phi X 174,
pubmed-meshheading:3038898-Cattle,
pubmed-meshheading:3038898-Cell Line,
pubmed-meshheading:3038898-Chromatography, Affinity,
pubmed-meshheading:3038898-Cricetinae,
pubmed-meshheading:3038898-DNA, Viral,
pubmed-meshheading:3038898-DNA Primase,
pubmed-meshheading:3038898-DNA Replication,
pubmed-meshheading:3038898-Drosophila,
pubmed-meshheading:3038898-Drug Stability,
pubmed-meshheading:3038898-Exonucleases,
pubmed-meshheading:3038898-Humans,
pubmed-meshheading:3038898-Immunologic Tests,
pubmed-meshheading:3038898-Lung,
pubmed-meshheading:3038898-Lymphocytes,
pubmed-meshheading:3038898-RNA Nucleotidyltransferases,
pubmed-meshheading:3038898-Thymus Gland
|
pubmed:year |
1987
|
pubmed:articleTitle |
On the fidelity of DNA replication. Isolation of high fidelity DNA polymerase-primase complexes by immunoaffinity chromatography.
|
pubmed:publicationType |
Journal Article,
Comparative Study,
Research Support, U.S. Gov't, P.H.S.
|