An application of protein engineering to determine the phase of crystal structure of Taq DNA polymerase

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Title
An application of protein engineering to determine the phase of crystal structure of Taq DNA polymerase
Author(s)
Dae Sil Lee
Bibliographic Citation
Journal of Biochemistry Molecular Biology & Biophysics, vol. 1, pp. 157-164
Publication Year
1998
Abstract
Taq DNA polymerase from Thermus aquaticus is used in the polymerase chain reaction. Not only is Taq DNA polymerase highly valuable in the commercial market for the polymerase chain reaction but also important in studying DNA replication. In order to determine the crystal structure of Taq DNA polymerase, it was necessary to use heavy atom multiple isomorphous replacement in addition to molecular replacement. The success of the multiple isomorphous replacement is often facilitated in a protein which has a reasonable number of cysteines; however, wild type Taw DNA polymerase has no cysteines. To provide binding sites for mercurys, three serines in Taq DNA polymerase were replaced by cysteines using site-directed mutagenesis. This allowed determination of the crystal structure of Taq DNA polymerase at 2.4 ? resolution by multiple isomorphous replacement.
Keyword
X-ray crystallographymolecular replacementmultiple isomorphous replacementPCRsite-directed mutagenesis
ISSN
I000-0095
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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