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Expression and Purification of the Human Xeroderma Pigmentosum F (XPF/ERCC1) Complex in Bacterial Cells for Protein Binding Assays and Crystallographic Studies
Author(s) -
Anduaga Javier,
Duprez Kevin,
Hilario Eduardo,
Fan Li
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.932.1
Subject(s) - xeroderma pigmentosum , nucleotide excision repair , cockayne syndrome , ercc1 , dna repair , complementation , biology , dna , recombinant dna , microbiology and biotechnology , endonuclease , genetics , transcription factor ii h , gene , mutant
Mutations in dna repair enzymes of the XP complementation group, XPA‐XPG, leads to a number of genetic disorders such as Xeroderma Pigmentosum, Cockayne's Syndrome, and Trichothiodystrophy. The structure specific dna repair endonuclease responsible for the 5′ incision during dna repair, XPF, is involved in homologous recombination that assists in removing interstrand cross‐link. XP‐F is an autosomal recessive disease characterized by hypersensitivity of the skin to sunlight followed by high incidence of skin cancer and frequent neurologic abnormalities. I expressed and purified human recombinant XPF and ERCC1 proteins in E.Coli Rosetta and BL21 cells, these results will be scaled up for use in interaction and crystallographic studies with proteins of the XP family.