K12polLF

DNA Polymerase I, Large (Klenow) Fragment
BBF10K_003248
source
Escherichia coli (strain K12)

DNA polymerase fragment that retains 5' → 3' polymerase activity and 3’ → 5’ exonuclease activity for removal of precoding nucleotides and proofreading, but loses 5' → 3' exonuclease activity. Usable for synthesis of double-stranded DNA from single-stranded templates, filling in of receded 3' ends of DNA fragments to make 5' overhang blunt, digesting away protruding 3' overhangs, preparation of radioactive DNA probes.

attr.
Chiara Gandini, Open Bioeconomy Lab

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-K12polLF
plasmid size
3944
insert size
1821
origin
ColE1 High Copy
copy number
high (3-500)
genbank file

Safety

BSL
BSL1

other information

No Value

References

primary references

NCBI
No Value
UniProt
P00582
Available Elsewhere
FALSE

protocols and instructions

https://docs.google.com/document/d/1c8PyT1Z9cWYUif4XQzcWl8MPbqVuL8-RNKCAancaVQU/edit, https://openbioeconomy.org/projects/expression-guide/

citations

Klenow, H., & Henningsen, I. (1970). Selective elimination of the exonuclease activity of the deoxyribonucleic acid polymerase from Escherichia coli B by limited proteolysis. Proceedings of the National Academy of Sciences, 65(1), 168-175. Joyce, C.M. and Grindley, N.D., 1983. Construction of a plasmid that overproduces the large proteolytic fragment (Klenow fragment) of DNA polymerase I of Escherichia coli. Proceedings of the National Academy of Sciences, 80(7), pp.1830-1834. https://doi.org/10.1073/pnas.80.7.1830 Ollis, D.L., Brick, P., Hamlin, R., Xuong, N.G. and Steitz, T.A., 1985. Structure of large fragment of Escherichia coli DNA polymerase I complexed with dTMP. nature, 313(6005), p.762.

intellectual property

We are unaware of third-party property rights claims on uses of this item.