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Site‐directed mutagenesis, expression, purification and translesion synthesis analysis of human DNA polymerase η mutations found in xeroderma pigmentosum variant and melanoma patients
Author(s) -
Kumar Mukesh,
Jiang Xiaohua
Publication year - 2013
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.27.1_supplement.976.2
Subject(s) - xeroderma pigmentosum , pyrimidine dimer , mutagenesis , dna polymerase , microbiology and biotechnology , point mutation , mutation , dna damage , dna , biology , polymerase , primer (cosmetics) , genetics , chemistry , gene , organic chemistry
The variant form of human syndrome Xeroderma Pigmentosum (XP‐V) is caused by mutations in DNA polymerase η (hpolη). XP‐V patients are deficient in repairing the DNA damage caused by ultraviolet (UV) light in the form of cyclobutane‐pyrimidine dimers (CPD). Many missense mutations in hpolη catalytic core (1–432aa) have been identified in XP‐V patients, and mutations have also been implicated in Melanoma. The aim of current research is to confirm that the catalytic core of hpolη is 1–432aa. Hpolη (1–432aa) has been constructed. It will be expressed in E.coli and tested in primer extension assays to assess its effectiveness during translesion synthesis (TLS). Several of the point mutations have been successfully introduced in to hpolη (1–432aa) by site‐directed mutagenesis. Those mutants are expressed in E.coli cells and will be examined for their activity during translesion synthesis (TLS). Source of research support: Student research grant to Mukesh Kumar, Department of Chemistry, TTU, Cookeville, TN, Faculty grant to Xiaohua Jiang, College of Arts and Sciences, TTU, Cookeville, TN