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Literature summary for 4.1.99.13 extracted from

  • Sancar, G.B.; Sancar, A.
    Purification and characterization of DNA photolyases (2006), Methods Enzymol., 408, 121-156.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
due to insolubility problems (6-4) photolyases is overexpressed as a fusion protein in Escherichia coli. Plasmid pXZ1997, a derivative of pMal-c2 containing the Drosophila melanogaster phr(6-4) cDNA fused in frame to the malE gene encoding maltose-binding protein (MBP), is propagated in Escherichia coli strain UNC523 (phr::kan uvrA::Tn10) selecting for ampicillin resistance. Cells are cultured in 2 liter of LB to A600: 0.6–0.8. IPTG is added to 0.3 mM, and incubation continued for 6 h prior to harvesting the cells by centrifugation. Drosophila melanogaster

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
110000
-
molecular weight of fusion protein, determined by SDS-PAGE Drosophila melanogaster

Organism

Organism UniProt Comment Textmining
Drosophila melanogaster
-
-
-

Purification (Commentary)

Purification (Comment) Organism
fusion protein is applied to amylose column. At this point the protein is above 90% pure. Further purification can be obtained by applying the eluted material to a 10 ml heparin-agarose column. Maltose-binding protein is removed by treatment with factor Xa protease Drosophila melanogaster

Storage Stability

Storage Stability Organism
glycerol to a final concentration of 50% (v/v) is added to the photolyase, storage at -70°C Drosophila melanogaster

Synonyms

Synonyms Comment Organism
(6-4) photolyase
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Drosophila melanogaster
phr (6-4)
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Drosophila melanogaster