Part:BBa_K5086004
Bst DNA Polymerase, Large Fragment
The large fragment of Bst DNA polymerase has strong thermal stability, strand displacement activity and polymerase activity, and is an essential enzyme in isothermal amplification technology. The optimal temperature for its catalysis is 45℃, and its catalytic isothermal nucleic acid amplification does not require thermal denaturation of DNA, which avoids the loss of time caused by the three-temperature cycle; at the same time, isothermal amplification has high sensitivity, and the detection limit is two orders of magnitude lower than that of the traditional PCR technology; its average amplification can reach 200 times, and there is a significant amplification band in 500-2000bp. It also has the advantages of high specificity, simple operation and easy to judge the results. In our molecular computing system, we mainly apply its 5'-3' polymerase active domain, which performs the catalytic polymerization function of the enzyme. The shape of the co-crystal structure formed when DNA polymerase combines with the primer-template is similar to that of the right hand of a human being, which can be divided into the palm region, the thumb region and the finger region. The palm region is the most conservative of the three structural domains, belonging to the catalytic site of the enzyme, and can catalyze the formation of nucleoside bonds; the thumb region determines the fidelity of DNA synthesis, and can make the newly synthesized double-stranded DNA move along the polymerase in a certain order, which is related to the shifting of the code mutation in the process of amplification; the finger region can selectively bind to the template strands of the dNTP complementary to the nucleotides, which is related to the specific binding of the nucleotide. At room temperature it catalyzes DNA polymerization using the target DNA as the primer. Since the polymerase fragment has strand displacement activity, the weighted strand that is base complementary paired with the template strand is gradually displaced from the template strand as the daughter strand DNA is synthesized.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 712
Illegal SpeI site found at 1468
Illegal PstI site found at 356 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 712
Illegal SpeI site found at 1468
Illegal PstI site found at 356 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 712
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 712
Illegal SpeI site found at 1468
Illegal PstI site found at 356 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 712
Illegal SpeI site found at 1468
Illegal PstI site found at 356 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 511
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