Application of N-terminally Truncated DNA Polymerase from Thermus thermophilus ( Tth Polymerase) to DNA Sequencing and Polymerase Chain Reactions: Comparative Study of Tth and Wild-Type Tth Polymerases
نویسندگان
چکیده
منابع مشابه
High-level overproduction of His-tagged Tth DNA polymerase in Thermus thermophilus.
A new plasmid for the overexpression of His-tagged thermozymes in Thermus thermophilus was developed. With this plasmid, soluble and active histidine-tagged DNA polymerase from T. thermophilus was overproduced in larger amounts in the thermophile than in Escherichia coli. The protein purified from the thermophile was active in PCR.
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DNA polymerase gene from Thermus aquaticus strain YT1 was amplified using VENTTM DNA po-lymerase and cloned under the control of X.PR promoter and expression was induced by a shift in tern perature. The culture was then sonicated, and after centrifugation the lysate was treated with polyethyleneimine followed by a salting-out step. Finally the protein was precipitated with ammonium sulfate and...
متن کاملAccuracy of replication in the polymerase chain reaction. Comparison between Thermotoga maritima DNA polymerase and Thermus aquaticus DNA polymerase.
For certain applications of the polymerase chain reaction (PCR), it may be necessary to consider the accuracy of replication. The breakthrough that made PCR user friendly was the commercialization of Thermus aquaticus (Taq) DNA polymerase, an enzyme that would survive the high temperatures needed for DNA denaturation. The development of enzymes with an inherent 3' to 5' exonuclease proofreading...
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ژورنال
عنوان ژورنال: DNA Research
سال: 1996
ISSN: 1340-2838
DOI: 10.1093/dnares/3.2.87