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Pfu DNA polymerase
''Pfu'' DNA polymerase is an enzyme found in the hyperthermophilic archaeon ''Pyrococcus furiosus'', where it functions to copy the organism's DNA during cell division. In the laboratory setting, ''Pfu'' is used to amplify DNA in the polymerase chain reaction (PCR), where the enzyme serves the central function of copying a new strand of DNA during each extension step.
==Proofreading ability of ''Pfu'' polymerase==
''Pfu'' DNA polymerase has superior thermostability and proofreading properties compared to ''Taq'' DNA polymerase. Unlike ''Taq'' DNA polymerase, ''Pfu'' DNA polymerase possesses 3' to 5' exonuclease proofreading activity, meaning that as the DNA is assembled from the 5' end to 3' end, the exonuclease activity immediately removes nucleotides misincorporated at the 3' end of the growing DNA strand. Consequently, ''Pfu'' DNA polymerase-generated PCR fragments will have fewer errors than ''Taq''-generated PCR inserts.
Commercially available ''Pfu'' typically results in an error rate of 1 in 1.3 million base pairs and can yield 2.6% mutated products when amplifying 1 kb fragments using PCR. However, ''Pfu'' is slower and typically requires 1–2 minutes per cycle to amplify 1kb of DNA at 72 °C. Using ''Pfu'' DNA polymerase in PCR reactions also results in blunt-ended PCR products.
''Pfu'' DNA polymerase is hence superior to ''Taq'' DNA polymerase for techniques that require high-fidelity DNA synthesis, but can also be used in conjunction with ''Taq'' polymerase to obtain the fidelity of ''Pfu'' with the speed of ''Taq'' polymerase activity..
抄文引用元・出典: フリー百科事典『 ウィキペディア（Wikipedia）』
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