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Overexpression of human polymerase ν in E. coli
Author(s) -
Shissler Susannah Claire,
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.976.5
Subject(s) - dna polymerase , processivity , subcloning , polymerase , biology , microbiology and biotechnology , dna replication , rna polymerase iii , dna polymerase ii , genetics , plasmid , dna , rna , reverse transcriptase , rna dependent rna polymerase , gene
The human genome has a vast repertoire of proteins involved in replication and repair of DNA. Among those proteins, there are 15 total polymerases, and polymerase ν (pol ν) is one of the least studied. It is a member of A‐family polymerases, which are highly conserved. Studies of human polymerase ν have described a significantly higher error rate due to lack of proof‐reading capabilities. Because of this, pol ν is highly regulated within the cell to prevent extraneous mutation. Although pol ν has been shown to participate in translesion synthesis in vitro, its precise function has yet to be determined. Our Research with pol ν will include subcloning the catalytic core into the vector pBG106. The plasmid will be transformed into E. coli cells and the protein will be over‐expressed. Histidine‐tagged pol ν should simplify the separation and purification of the protein by use of a Nickel affinity column.