topA
DNA topoisomerase 1BBF10K_001044
source
Escherichia coli str. K-12 substr. MG1655
Releases the supercoiling and torsional tension of DNA, which is introduced during the DNA replication and transcription, by transiently cleaving and rejoining one strand of the DNA duplex. Introduces a single-strand break via transesterification at a target site in duplex DNA. The scissile phosphodiester is attacked by the catalytic tyrosine of the enzyme, resulting in the formation of a DNA-(5'-phosphotyrosyl)-enzyme intermediate and the expulsion of a 3'-OH DNA strand. The free DNA strand then undergoes passage around the unbroken strand, thus removing DNA supercoils. Finally, in the religation step, the DNA 3'-OH attacks the covalent intermediate to expel the active-site tyrosine and restore the DNA phosphodiester backbone.
attr.
Keoni Gandall
Usage
growth
shipping strain
{shipping_strain}
growth conditions
37 C, shaking 300 rpm
antibiotic
ampicillin
expression
strain
N/A
promoter
N/A
inducer
N/A
cloning
method
GoldenGate
enzyme
BsaI
overhangs
3' - AATG … GCTT - 5'
sequencing
forward primer
M13 For
reverse primer
M13 Rev
Construct
plasmid name
pOpen-topA
plasmid size
4721
insert size
2598
origin
ColE1 High Copy
copy number
500-700
Safety
BSL
BSL1
other information
No ValueReferences
Available Elsewhere
FALSE