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KMID : 0578319950050050452
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Molecules and Cells
1995 Volume.5 No. 5 p.452 ~ p.460
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An Application of Genetic Engineering for Solving the Crystal Structure of Taq DNA Polymerase
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Kim Youngsoo
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Abstract
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Taq DNA polymerase from Thermus aquaticus has been shown to be very useful in the polymerase chain reaction (PCR). Not only is Taq DNA polymerase highly valuable in the commercial market for PCR, but it is also important in studying DNA replication. Taq DNA polymerase is apparently homologous to E. coli DNA polymerase I. The crystal structure of Taq DNA polymerase could be useful for both PCR application and DNA replication studies. Crystal structure in the form of electron density may be represented as the sum of X-ray diffraction intensity and phases from crystal. The phase information can be generally obtained from multiple isomorphous replacement (MIR) in X-ray crystallography. The success of multiple isomorphous replacement largely depends on having a reasonable number of cysteines in the protein, but Taq DNA polymerase does not carry any at all. The three serines in Taq DNA polymerase were substituted with cysteines by site-directed mutagenesis, which eventually brought out appropriate phasing information by MIR to determine the crystal structure of Taq DNA polymerase to 2.4 Å resolution.
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